Sialyl-Lewisa and sialyl-Lewisx epitope analogues

ABSTRACT

Sialyl-Lewis x  and sialyl-Lewis a  epitope analogues in which the naturally occurring N-acetyl group of the N-acetylglucosamine monomer is replaced by various aliphatic or aromatic substituents and the L-fucose naturally present is replaced by various naturally occurring or non-naturally occurring sugars.

This application is the U.S. national stage entry under 35 U.S.C. §371of PCT/EP97/00222, filed Jan. 17, 1997.

The invention relates to sialyl-Lewis^(a) and sialyl-Lewis^(x) epitopeanalogues, their preparation and use, and compositions comprising thesecompounds.

Carbohydrate domains and cell surfaces play a role in the treatment ofmany diseases, for example viral and bacterial infections, inflammatorydiseases, rheumatic arthritis, allergies, post-infarction syndromes,septic shock, apoplexy, acute and chronic organ rejections, sepsis andcancer (formation of metastases) [Witczak, Z. J., Current Med. Commun.1:392-405 (1995)]. Carbohydrate epitopes on eukaryotic cells are used byviruses, bacteria and toxins as specific adhesion points [Edwards, M.,Curr. Op. in Therapeutic Patents 1617-1630 (1991)]. Carbohydrate domainsalso function as receptors of roaming malignant cells [Muramatsu, T.,Glycobiology 3:294-296 (1993)]. However, they are also specific bindingepitopes for certain transmembrane proteins, for example E-, P- andL-selectins. Selectins are found in the surface of both endothelialcells and circulating cells of the haemato-lymphoid system. They undergospecific interactions with carbohydrates [Lasky, L. A., Ann. Rev.Biochem. 64:113-139 (1995); Nelson, R. M., Dolich, S., Aruffo, A.,Cecconi, O., Bevilacqua, M. P., J. Clin. Invest. 91:1157-1166 (1993)].

Sialylated and/or fucosylated carbohydrate epitopes are chiefly heldresponsible for such adhesion phenomena [Varki, A., Glycobiology3:97-130 (1993)]. The two tetrasaccharide epitopes sialyl-Lewis^(a)[αsia(2→3)βgal(1→3)[αfuc(1→4)]-βglcNAc-OR] and sialyl-Lewis^(x)[αsia(2→3)βgal(1→4)[αfuc(1→3)]-βglcNAc-OR] (in which R must be analgycon having at least one carbon atom) are attributed particularimportance in pathogenic inflammatory processes [Fukuda, M., Bioorg.Med. Chem. 3:207-215 (1995)].

Several routes have already been taken to isolate derivatives of thesecarbohydrate epitopes with better binding affinities than the naturallyoccurring ligand and an increased physiological stability. On the onehand, the natural epitope has been modified only slightly. Thus,N-acetylglucosamine has been replaced by sugars, such as glucosamine orglucose (WO 93/10,796), or by straight-chain or cyclic aliphaticradicals (EP 671,408). On the other hand, as many of the sugar monomersof the epitope as possible have been replaced by other functional units[Allanson, N. M., Davidson, A. H., Floyd, C. D., Martin, F. M.,Tetra-hedron Assym. 5:2061-2076 (1994)]. However, none of these variousapproaches has so far led to epitope analogues having a significantlyhigher binding affinity. WO 94/26,760 discloses that compounds havinghigher binding affinities for selectins can be obtained if the N-acetylgroup of N-acetylglucosamine, which is regarded as a group which is notrelevant to binding (EP 671,408), is replaced by aromatic amides.

Surprisingly, the present invention provides sialyl-Lewis^(x) andsialyl-Lewis^(a) epitope analogues having an improved binding affinityfor the corresponding selecting, in which the naturally occurringN-acetyl group of the N-acetylglucosamine monomer is replaced by variousaliphatic and aromatic substituents and the L-fucose naturally presentis replaced by various naturally occurring and non-naturally occurringsugars.

The present invention relates to compounds of the formula I or II

in which Z is an α-bonded pyranose of the formula III

with the proviso that Z is not L-fucose,

R₁ is hydrogen, C₁-C₂₀alkyl, C₁-C₂₀alkenyl, C₃-C₁₅cycloalkyl or a mono-or bicyclic C₆-C₁₀aryl or C₂-C₉heteroaryl, where alkyl, alkenyl,cycloalkyl, aryl and heteroaryl are unsubstituted or mono- orpolysubstituted by a substituent chosen from the group consisting of OH,halogen, halo-C₁-C₁₈alkyl, nitro, C₁-C₁₈alkyl, C₁-C₁₈alkoxy, amino,mono-C₁-C₁₈alkylamino, di-C₁-C₁₈alkylamino, benzylamino, sulfhydryl,thio-C₁-C₁₈alkyl and C₁-C₁₈alkylcarboxamide;

R₂ is C₁-C₁₈alkyl, mono- or polysubstituted C₁-C₁₈alkyl, C₃-C₈cycloalkylor mono- or polysubstituted C₃-C₈cycloalkyl, where one or more CH₂groups in the alkyl and in the cycloalkyl, where appropriate,independently of one another are replaced by oxygen, sulfur or an iminogroup and the substituents are chosen from the group consisting of OH,SH, NH₂, carboxamide, C(O)O and C₁-C₁₈alkoxycarbonyl;

R₃ is a methyl or hydroxymethyl group;

the individual R₄ independently of one another are hydrogen, OH,C₁-C₈alkyl, O-C₁-C₈alkyl, halogen, NH₂, SH or NHC(O)-C₁-C₈alkyl;

R₅ is hydrogen, C₁-C₈alkyl or (CH₂)_(m)R₄, in which m is a number from 1to 5; and

X is —C(O)—, —C(S)—, —S(O)₂—, —C(O)Y— or —C(S)Y—, in which

Y is NH, O, S, S-C₁-C₆alkylene, NH-C₁-C₆alkylene or O-C₁-C₆alkylene.

In the context of the present invention, the pyranose is advantageouslyD-fucose, D,L-arabinose, D,L-ribose, D,L-xylose, D,L-lyxose, L-rhamnose,D,L-galactose, D,L-glucose, D,L-mannose, D,L-gulose, D,L-allose,D,L-altrose, D,L-idose or D,L-talose, in particular D-fucose,D-arabinose, L-galactose or L-glucose. Preferred compounds are those inwhich the pyranose is an α-bonded D-fucose, D-arabinose, L-galactose orL-glucose, in which one or more R₄ independently of one another arehydrogen, halogen, sulfhydryl, a thioalkyl group, an amino group, anaminoalkyl group, a dialkylamino group or an aminoacyl group; and wherethe alkyl, where appropriate independently of one another, is a linearor branched C₁-C₁₈alkyl.

In the context of the present invention, the aryl or heteroaryl is afive- or six-membered ring or a bicyclic radical of two fused six- orfive-membered rings or one six-membered and one five-membered ring, oneor more heteroatoms chosen from the group consisting of the oxygen,nitrogen and sulfur atom being present in the heteroaryl. Examples arederived from benzene, pentalene, naphthalene, indene, furan, pyrrole,pyrazole, imidazole, isoxazole, oxazole, furazan, thiadiazole,thiophene, thiazole, oxadiazole, triazole, indole, indazole, purine,benzimidazole, benzoxazole, benzothiazole, pyran, pyridine, pyridazine,triazine, pyrimidine, pyrazine, isoquinoline, cinnoline, phthalazine,quinoline, quinazoline, pteridine, benzotriazine or quinoxaline.

Halogen is preferably F, Cl or Br.

The abovementioned alkyl and alkylene can be linear or branched. Someexamples of alkyl, alkoxy, thioalkyl and alkylamino, which preferablycontain 1 to 12 C atoms, are methyl, ethyl and the isomers of propyl,butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl and dodecyl,and corresponding alkoxy, thioalkyl and alkylamino radicals. Preferredalkyl, alkoxy, thioalkyl and alkylamino radicals are methyl, ethyl, n-and i-propyl, n-, i- and t-butyl, methoxy, ethoxy, isopropyloxy,methylthio, isopropylthio and ethylthio, aminomethyl, aminoisopropyl andaminoethyl.

Examples of alkenyl are allyl, but-1-en-3- or -4-yl, pent-3- or-4-en-1-, -2- or -3-yl, hex-3-, -4- or -5-en-1- or -2-yl and(C₁-C₄alkyl)CH═CH—CH₂—. Examples of cycloalkyl are cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl.

In the context of the present invention, preferred compounds of theformula I or II are those in which R₁ is hydrogen, C₁-C₂₀alkyl orC₁-C₂₀alkenyl, which are unsubstituted or mono- or polysubstituted by asubstituent chosen from the group consisting of OH, halogen,halo-C₁-C₁₈alkyl, nitro, C₁-C₁₈alkyl, C₁-C₁₈alkoxy, amino,mono-C₁-C₁₈alkylamino, di-C₁-C₁₈alkyl-amino, benzylamino, sulfhydryl,thio-C₁-C₁₈alkyl and C₁-C₁₈alkylcarboxamide. Particularly preferredcompounds are those in which R₁ is C₁-C₁₀alkyl or C₁-C₁₀alkenyl, whichare unsubstituted or mono- or polysubstituted by a substituent chosenfrom the group consisting of OH, halogen, halo-C₁-C₁₈alkyl, nitro,C₁-C₁₈alkyl, C₁-C₁₈alkoxy, amino, mono-C₁-C₁₈alkylamino,di-C₁-C₁₈alkylamino, benzylamino, sulfhydryl, thio-C₁-C₁₈alkyl andC₁-C₁₈alkylcarboxamide. Particularly preferred compounds are those inwhich R₁ is C₁-C₅alkyl or C₁-C₅alkenyl, which are unsubstituted orsubstituted by OH or halogen, —CH₃, —CF₃, —CH₂—CH═CH₂, —CH₂OH and —CH₂SHbeing especially preferred.

Compounds of the formula I or II which are furthermore preferred arethose in which R₁ is a mono- or bicyclic C₆-C₁₀aryl or C₂-C₉heteroaryl,which are unsubstituted or mono- or polysubstituted by a substituentchosen from the group consisting of OH, halogen, halo-C₁-C₁₈alkyl,nitro, C₁-C₁₈alkyl, C₁-C₁₈alkoxy, amino, mono-C₁-C₁₈alkylamino,di-C₁-C₁₈alkylamino, benzylamino, sulfhydryl, thio-C₁-C₁₈alkyl andC₁-C₁₈alkylcarboxamide. Particularly preferred compounds of the formulaI or II are those in which R₁ is a mono- or bicyclic C₆-C₁₀aryl orC₂-C₉heteroaryl, which are substituted by at least one OH and are notfurther substituted or are further mono- or polysubstituted by asubstituent chosen from the group consisting of halogen,halo-C₁-C₁₈alkyl, nitro, C₁-C₁₈alkyl, C₁-C₁₈alkoxy, amino,mono-C₁-C₁₈alkylamino, di-C₁-C₁₈alkylamino, benzylamino, sulfhydryl,thio-C₁-C₁₈alkyl and C₁-C₁₈alkylcarboxamide. Especially preferredcompounds of the formula I or II are those in which R₁ is phenyl or amono- or bicyclic C₄-C₉heteroaryl, which are substituted by at least oneOH and are not further substituted or are further substituted by asubstituent chosen from the group consisting of halogen, nitro,C₁-C₁₈alkyl and C₁-C₁₈alkoxy. Very particularly preferred compounds arethose in which R₁ is phenyl, which is substituted by one OH and F, NO₂,CH₃ or OCH₃ or by two OH; or in which R₁ is a C₄heteroaryl which issubstituted by two OH, or a C₉heteroaryl which is substituted by one OH.

In the context of the present invention, preferred compounds of theformula I or II are furthermore those in which R₂ is C₁-C₁₈alkyl, mono-or polysubstituted C₁-C₁₈alkyl, C₃-C₈cycloalkyl or mono- orpolysubstituted C₃-C₈cycloalkyl, where the substituents are chosen fromthe group consisting of OH, SH, NH₂, carboxamide, C(O)O andC₁-C₁₈alkoxycarbonyl. R₂ is particularly preferably C₁-C₁₈alkyl orC₁-C₁₈alkyl which is mono- or polysubstituted independently of oneanother by OH, SH, NH₂, carboxamide, C(O)O or C₁-C₁₈alkoxycarbonyl, R₂is especially preferably C₁-C₁₈alkyl or C₁-C₁₈alkyl monosubstituted byC(O)O, and R₂ is most preferably —(CH₂)₈COOCH₃.

In the context of the present invention, compounds of the formula I orII which are preferred are furthermore those in which R₃ is methyl.

Moreover, compounds of the formula I or II which are preferred are thosein which the individual R₄ independently of one another are hydrogen,OH, C₁-C₄alkyl, O-C₁-C₄alkyl, halogen, NH₂ or NHC(O)-C₁-C₈alkyl.Particularly preferred compounds are those in which the individual R₄independently of one another are OH, halogen or NH₂, especially those inwhich all the R₄ are OH or two R₄ are OH and one R₄ is halogen, inparticular F, or NH₂.

Preferred compounds of the formula I or II are those in which R₅ ishydrogen, C₁-C₈alkyl or (CH₂)_(m)OH, in which m is an integer from 1 to5, particularly preferably H, C₁-C₄alkyl or (CH₂)_(m)OH, in which m is 1or 2, especially preferably hydrogen, CH₃ or CH₂OH.

In preferred compounds of the formula I or II, X is —C(O)—, —S(O)₂— or—C(O)Y—, in which Y is —NH—, —S-C₁-C₆-alkylene or —O-C₁-C₆alkylene, andX is, in particular, —C(O)—, —S(O)₂—, —C(O)SCH₂ or —C(O)OCH₂.

Preferred compounds of the formula I or II are, in particular, those inwhich

R₁ is hydrogen, C₁-C₂₀alkyl or C₁-C₂₀alkenyl, which are unsubstituted ormono- or polysubstituted by a substituent chosen from the groupconsisting of OH, halogen, halo-C₁-C₁₈alkyl, nitro, C₁-C₁₈alkyl,C₁-C₁₈alkoxy, amino, mono-C₁-C₁₈alkylamino, di-C₁-C₁₈alkylamino,benzylamino, sulfhydryl, thio-C₁-C₁₈alkyl and C₁-C₁₈alkylcarboxamide;

R₂ is C₁-C₁₈alkyl, mono- or polysubstituted C₁-C₁₈alkyl, C₃-C₈cycloalkylor mono- or polysubstituted C₃-C₈cycloalkyl, where the substituents arechosen from the group consisting of OH, SH, NH₂, carboxamide, C(O)O andC₁-C₁₈alkoxycarbonyl;

R₃ is methyl;

the individual R₄ independently of one another are hydrogen, OH,C₁-C₄alkyl, O-C₁-C₄alkyl, halogen, NH₂ or NHC(O)-C₁-C₈alkyl;

R₅ is hydrogen, C₁-C₈alkyl or (CH₂)_(m)OH, in which m is a number from 1to 5; and

X is —C(O)—, —S(O)₂— or —C(O)Y—, in which

Y is —NH—, —S-C₁-C₆alkylene or —O-C₁-C₆alkylene.

Very particularly preferred compounds of the formula I or II are thosein which

R₁ is C₁-C₁₀alkyl or C₁-C₁₀alkenyl, which are unsubstituted or mono- orpolysubstituted by a substituent chosen from the group consisting of OH,halogen, halo-C₁-C₁₈alkyl, nitro, C₁-C₁₈alkyl, C₁-C₁₈alkoxy, amino,mono-C₁-C₁₈alkylamino, di-C₁-C₁₈alkylamino, benzylamino, sulfhydryl,thio-C₁-C₁₈alkyl and C₁-C₁₈alkylcarboxamide;

R₂ is C₁-C₁₈alkyl or C₁-C₁₈alkyl which is mono- or polysubstitutedindependently of one another by OH, SH, NH₂, carboxamide, C(O)O orC₁-C₁₈alkoxycarbonyl;

R₃ is methyl;

the individual R₄ independently of one another are OH, halogen or NH₂;

R₅ is H, C₁-C₄alkyl or (CH₂)_(m)OH, in which m is 1 or 2; and

X is —C(O)—, —S(O)₂—, —C(O)SCH₂ or —C(O)OCH₂.

Of these compounds, especially preferred compounds are those in which

R₁ is C₁-C₅alkyl or C₁-C₅alkenyl, which are unsubstituted or substitutedby OH or halogen;

R₂ is C₁-C₁₈alkyl or C₁-C₁₈alkyl which is monosubstituted by C(O)O;

all the R₄ are OH or two R₄ are OH and one R₄ is halogen or NH₂; and

R₅ is hydrogen, CH₃ or CH₂OH.

Especially preferred compounds within this group are those in which R₁is —CH₃, —CH₂—CH═CH₂, —CF₃ or —CH₂OH; R₂ is —(CH₂)₈COOCH₃; all the R₄are OH or two R₄ are OH and one R₄ is F or NH₂; and R₅ is hydrogen, CH₃or CH₂OH.

Preferred compounds of the formula I or II are furthermore, inparticular, those in which

R₁ is a mono- or bicyclic C₆-C₁₀aryl or C₂-C₉heteroaryl, which areunsubstituted or mono- or polysubstituted by a substituent chosen fromthe group consisting of OH, halogen, halo-C₁-C₁₈alkyl, nitro,C₁-C₁₈alkyl, C₁-C₁₈alkoxy, amino, mono-C₁-C₁₈alkylamino,di-C₁-C₁₈alkylamino, benzylamino, sulfhydryl, thio-C₁-C₁₈alkyl andC₁-C₁₈alkylcarboxamide;

R₂ is C₁-C₁₈alkyl, mono- or polysubstituted C₁-C₁₈alkyl, C₃-C₈cycloalkylor mono- or polysubstituted C₃-C₈cycloalkyl, where the substituents arechosen from the group consisting of OH, SH, NH₂, carboxamide, C(O)O andC₁-C₁₈alkoxycarbonyl;

R₃ is methyl;

the individual R₄ independently of one another are hydrogen, OH,C₁-C₄alkyl, O-C₁-C₄alkyl, halogen, NH₂ or NHC(O)-C₁-C₈alkyl;

R₅ is hydrogen, C₁-C₈alkyl or (CH₂)_(m)OH, in which m is a number from 1to 5; and

X is —C(O)—, —S(O)₂— or —C(O)Y—, in which

Y is —NH—, —S-C₁-C₆alkylene or —O-C₁-C₆alkylene.

Very particularly preferred compounds of the formula I or II are thosein which

R₁ is a mono- or bicyclic C₆-C₁₀aryl or C₂-C₉heteroaryl, which aresubstituted by at least one OH and are not further substituted or arefurther mono- or polysubstituted by a substituent chosen from the groupconsisting of halogen, halo-C₁-C₁₈alkyl, nitro, C₁-C₁₈alkyl,C₁-C₁₈alkoxy, amino, mono-C₁-C₁₈alkylamino, di-C₁-C₁₈alkylamino,benzylamino, sulfhydryl, thio-C₁-C₁₈alkyl and C₁-C₁₈alkylcarboxamide;

R₂ is C₁-C₁₈alkyl or C₁-C₁₈alkyl which is mono- or polysubstitutedindependently of one another by OH, SH, NH₂, carboxamide, C(O)O orC₁-C₁₈alkoxycarbonyl;

R₃ is methyl;

the individual R₄ independently of one another are OH, halogen or NH₂;

R₅ is H, C₁-C₄alkyl or (CH₂)_(m)OH, in which m is 1 or 2; and

X is —C(O)—, —S(O)₂—, —C(O)SCH₂ or —C(O)OCH₂.

Of these compounds, especially preferred compounds are those in which

R₁ is phenyl or a mono- or bicyclic C₄-C₉heteroaryl, which aresubstituted by at least one OH and are not further substituted or arefurther substituted by a substituent chosen from the group consisting ofhalogen, nitro, C₁-C₁₈alkyl and C₁-C₁₈alkoxy;

R₂ is C₁-C₁₈alkyl or C₁-C₁₈alkyl which is monosubstituted by C(O)O;

all the R₄ are OH or two R₄ are OH and one R₄ is halogen or NH₂; and

R₅ is hydrogen, CH₃ or CH₂OH.

Within this group, especially preferred compounds are those in which R₁is phenyl, which is substituted by one OH and F, NO₂, CH₃ or OCH₃ or bytwo OH; or in which R₁ is a C₄heteroaryl which is substituted by two OH,or a C₉heteroaryl which is substituted by one OH; R₂ is —(CH₂)₈COOCH₃;all the R₄ are OH or two R₄ are OH and one R₄ is F or NH₂; and R₅ ishydrogen, CH₃ or CH₂OH.

The most preferred compounds of the formula I are those in which R₂ is—(CH₂)₈COOCH₃; R₃ is methyl; and

(a) R₁ is hydrogen; Z is an α-bonded L-galactose; and X is —C(O)—;

(b) R₁ is —CH₂—CH═CH₂; Z is an α-bonded L-galactose; and X is—C(O)OCH₂—;

(c) R₁ is —CH₂—CH═CH₂; Z is an α-bonded D-arabinose; and X is—C(O)OCH₂—;

(d) R₁ is 4-hydroxy-3-methoxy-phenyl; Z is an α-bonded D-arabinose; andX is —C(O)—;

(e) R₁ is 4-hydroxy-3-methoxy-phenyl; Z is an α-bonded L-galactose; andX is —C(O)—;

(f) R₁ is 2-hydroxy-5-methyl-phenyl; Z is an α-bonded D-arabinose; and Xis —C(O)—;

(g) R₁ is 2-hydroxy-5-methyl-phenyl; Z is an α-bonded L-galactose; and Xis —C(O)—;

(h) R₁ is 2-hydroxy-3-nitro-phenyl; Z is an α-bonded L-galactose; and Xis —C(O)—;

(i) R₁ is 2-hydroxy-5-fluoro-phenyl; Z is an α-bonded D-arabinose; and Xis —C(O)—;

(j) R₁ is 3,5-dihydroxy-phenyl; Z is an α-bonded D-arabinose; and X is—C(O)—;

(k) R₁ is 3,5-dihydroxy-phenyl; Z is an α-bonded L-galactose; and X is—C(O)—;

(l) R₁ is 3,5-dihydroxy-pyrimidinyl; Z is an α-bonded D-arabinose; and Xis —C(O)—;

(m) R₁ is 3,5-dihydroxy-pyrimidinyl; Z is an α-bonded L-galactose; and Xis —C(O)—; or

(n) R₁ is 2-(8-hydroxy)quinolinyl; Z is an α-bonded L-galactose; and Xis —C(O)—.

Within this group, particularly preferred compounds of the formula I arethose in which R₂ is —(CH₂)₈COOCH₃; R₃ is methyl; Z is an α-bondedL-galactose; X is —C(O)— and R₁ is hydrogen; 4-hydroxy-3-methoxy-phenyl;2-hydroxy-5-methyl-phenyl; 2-hydroxy-3-nitrophenyl;3,5-dihydroxy-phenyl; 3,5-dihydroxy-pyrimidinyl or2-(8-hydroxy)quinolinyl. That compound in which R₂ is —(CH₂)₈COOCH₃; R₃is methyl; Z is an α-bonded L-galactose; X is —C(O)— and R₁ is4-hydroxy-3-methoxy-phenyl is especially preferred.

The most preferred compounds of the formula II are those in which R₂ is—(CH₂)₈COOCH₃; R₃ is methyl; and

(a) R₁ is hydrogen; Z is an α-bonded D-arabinose; and X is —C(O)—;

(b) R₁ is hydrogen; Z is an α-bonded L-2-fluoro-fucose; and X is —C(O)—;

(c) R₁ is CH₃; Z is an α-bonded D-arabinose; and X is —C(O)—;

(d) R₁ is CH₃; Z is an α-bonded L-2-fluoro-fucose; and X is —C(O)—;

(e) R₁ is CH₃; Z is an α-bonded L-2-aminoiucose; and X is —C(O)—;

(f) R₁ is CH₃; Z is an α-bonded L-galactose; and X is —C(O)—;

(g) R₁ is CH₃; Z is an α-bonded L-glucose; and X is —C(O)—;

(h) R₁ is CH₃; Z is an α-bonded L-galactose; and X is —C(O)OCH₂—;

(i) R₁ is CH₃; Z is an α-bonded L-glucose; and X is —C(O)OCH₂—;

(j) R₁ is CH₃; Z is an α-bonded D-arabinose; and X is S(O)₂;

(k) R₁ is CH₃; Z is an α-bonded D-arabinose; and X is —C(O)SCH₂—;

(l) R₁ is CF₃; Z is an α-bonded D-arabinose; and X is —C(O)—;

(m) R₁ is CH₂OH; Z is an α-bonded D-arabinose; and X is —C(O)—;

(n) R₁ is —CH₂—CH═CH₂; Z is an α-bonded D-arabinose; and X is—C(O)OCH₂—;

(o) R₁ is —CH₂—CH═CH₂; Z is an α-bonded L-galactose; and X is—C(O)OCH₂—;

(p) R₁ is phenyl; Z is an α-bonded L-galactose; and X is —C(O)OCH₂—;

(q) R₁ is 2-hydroxy-5-methyl-phenyl; Z is an α-bonded D-arabinose; and Xis —C(O)—;

(r) R₁ is 2-hydroxy-5-methyl-phenyl; Z is an α-bonded L-galactose; and Xis —C(O)—;

(s) R₁ is 2-hydroxy-5-mluoro-phenyl; Z is an α-bonded D-arabinose; and Xis —C(O)—;

(t) R₁ is 4-hydroxy-3-methoxy-phenyl; Z is an α-bonded D-arabinose; andX is —C(O)—;

(u) R₁ is 3,5-dihydroxy-phenyl; Z is an α-bonded L-galactose; and X is—C(O)—;

(v) R₁ is 3,5-dihydroxy-phenyl; Z is an α-bonded L-2-amino-fucose; and Xis —C(O)—;

(w) R₁ is 3,5-dihydroxy-phenyl; Z is an α-bonded D-arabinose; and X is—C(O)OCH₂— or

(x) R₁ is 3,5-dihydroxy-pyrimidinyl; Z is an α-bonded D-arabinose; and Xis —C(O)—.

Within this group, particularly preferred compounds of the formula IIare those in which R₁ is CH₃; R₂ is —(CH₂)₈COOCH₃; R₃ is methyl; Z is anα-bonded L-galactose and X is —C(O)— or —C(O)OCH₂—.

The present invention furthermore relates to a process for thepreparation of compounds of the formula I, which comprises

(a) reacting a compound of the formula V

R₇—X′—R₁  (V),

 in which

(a′) R₇ is halogen, X′ is as defined above for X and R₁ is as definedabove, or

(a″) R₇ is C(O) or C(S), X′ is —N═ and R₁ is as defined above, or

(a′″) R₇ is OH, X′ is as defined above for X and R₁ is as defined above,directly after in situ activation analogously to methods such as arecustomary in peptide chemistry [Bodansky, M., Principles of PeptideChemistry, 2nd Edition 16-61, Springer Berlin (1993)], with a compoundof the formula IV

 in which R₂ is as defined above and the individual R₄ independently ofone another are hydrogen, acetyl, propionyl, butyryl or benzoyl,

any acetyl, propionyl, butyryl or benzoyl groups present being split offwith a basic alcohol solution,

to give a compound of the formula VI

 in which R₂, R₁ and X are as defined above;

(b) reacting the compound of the formula VI with uridinedi-phosphate-galactose in the presence of β(1→4)galactose transferaseand then with cytidine mono-phosphate-sialic acid in the presence ofα(2→3)sialic acid transferase to give a compound of the formula VII

 in which R₁, R₂, R₃ and X are as defined above, and

(c) reacting the resulting product with a guanosinedi-phosphate-activated donor of the formula XI

 in which R₄ and R₅ are as defined above, in the presence of fucosetransferase VI to give a compound of the formula I.

The present invention furthermore relates to a process for thepreparation of compounds of the formula II, which comprises

(a) reacting a compound of the formula VI with uridinedi-phosphate-galactose in the presence of β(1→4)galactose transferaseand then with cytidine mono-phosphate-sialic acid in the presence ofα(2→3)sialic acid transferase to give a compound of the formula VII and

(b) reacting the resulting product with a compound of the formula XI inthe presence of fucose transferase to give a compound of the formula I.

The present invention furthermore relates to a process for thepreparation of compounds of the formula II which comprises

(a) reacting a compound of the formula IV with a compound of the formulaV as described for the preparation of the compounds of the formula I,

(b) reacting the compound of the formula VI with uridinedi-phosphate-galactose in the presence of β(1→3)galactose transferaseand then cytidine mono-phosphate-sialic acid in the presence ofα(2→3)sialic acid transferase to give a compound of the formula VIII

 in which R₁, R₂, R₃ and X are as defined above, and

(c) reacting the resulting product with a compound of the formula XI inthe presence of fucose transferase to give a compound of the formula II.

The present invention furthermore relates to a process for thepreparation of compounds of the formula II, which comprises

(a) reacting a compound of the formula VI with uridinedi-phosphate-galactose in the presence of β(1→3)galactose transferaseand then with cytidine mono-phosphate-sialic acid in the presence ofα(2→3)sialic acid transferase to give a compound of the formula VIII and

(b) reacting the resulting product with a compound of the formula XI inthe presence of fucose transferase to give a compound of the formula II.

The present invention furthermore relates to a process for thepreparation of compounds of the formula II, which comprises

(a) reacting a compound of the formula V

R₇—X′—R₁  (V),

 in which

(a′) R₇ is halogen, X′ is as defined above for X and R₁ is as definedabove, or

(a″) R₇ is C(O) or C(S), X′ is —N═ and R₁ is as defined above, or

(a′″) R₇ is OH, X′ is as defined above for X and R₁ is as defined above,directly after in situ activation analogously to methods such as arecustomary in peptide chemistry [Bodansky, M., Principles of PeptideChemistry, 2nd Edition, 16-61, Springer Berlin (1993)], with a compoundof the formula IX

 in which R₂ is as defined above and the individual R₄ independently ofone another are hydrogen, acetyl, propionyl, butyryl or benzoyl,

any acetyl, propionyl, butyryl or benzoyl groups present being split offwith a basic alcohol solution,

to give a compound of the formula X

 in which R₂, R₁ and X are as defined above;

(b) reacting the compound of the formula X with cytidinemono-phosphate-sialic acid in the presence of α(2→3)sialic acidtransferase to give a compound of the formula VIII

 in which R₁, R₂, R₃ and X are as defined above, and

(c) reacting the resulting product with a compound of the formula XI inthe presence of fucose transferase to give a compound of the formula II.

The present invention furthermore relates to a process for thepreparation of compounds of the formula II, which comprises

(a) reacting a compound of the formula X with cytidinemono-phosphate-sialic acid in the presence of α(2→3)sialic acidtransferase to give a compound of the formula VIII and

(b) reacting the resulting product with a compound of the formula XI inthe presence of fucose transferase to give a compound of the formula II.

With the enzymatic process according to the invention, it is possiblefor oligosaccharide structures to be prepared more efficiently comparedwith the chemical syntheses to date, and for highly modified,non-naturally occurring substrates to be glycosylated enzymatically in ahighly regio- and stereoselective manner, it being possible for thecompounds according to the invention to be prepared without the use ofhighly toxic heavy metal promoters (for example Hg²⁺ salts), such as areusually employed in chemical glycosylations.

The compounds of the formulae IV, V and IX are known or can be preparedby known processes. The compounds of the formula IX can be synthesizedby a process of Lemieux et al. and Boullanger et al. [Lemieux, R. U.,Bundle, D. R., Baker, D. A., J. Am. Chem. Soc. 97:4076-4083 (1975);Boullanger, P., Banoub, J., Descotes, G., Can. J. Chem. 65:1343-1348(1987)].

The amidation of compounds of the formulae IV and IX can be carried outin various way, depending on the definition of R₁, R₇ and X [Bodansky,M., Principles of Peptide Chemistry, 2nd Edition, 9-62, Springer Berlin(1993)].

For example, in case (a), R₇ is OH and X and R₁ are as defined above,the amidation can be carried out directly after the compounds of theformula V have first been activated with a diimidazole, for examplecarbonyldiimidazole (CDI), in a polar non-protic solvent, such asdimethylformamide (DMF) or acetonitrile.

(b) The amidation in the case of these compounds of the formulae IV andIX can also first be carried out after the aromatic OH groups haveinitially been protected, for example acetylated or benzoylated[McCorkindale, N. J., Roy, T. P., Hutchinson, S. A., Tetrahedron2:1107-1111 (1972)]. The acid function can then be converted into theacid chloride with an inorganic acid chloride, for example thionylchloride. These products are then coupled with the amine of the formulaIV or IX in the presence of a base, for example triethylamine, in asolvent, such as methylene chloride, and the products are converted intothe glucosamide derivatives of the formula VI or X by addition of abasic alcohol solution, for example methanol solution.

(c) Chlorides of the formula V which can undergo coupling, where R₇ isCl, X is C(O)-C₁-C₆alkylene and R₁ is as defined above, are obtained byacetylating the aromatic OH groups of the corresponding carboxylic acidand initially reducing the free acid function to the benzylic OH groupsby means of diborane [McCorkindale, N. J., Roy, T. P., Hutchinson, S.A., Tetrahedron 2:1107-1111 (1972)]. This product is reacted withphosgene to give the corresponding alkoxycarbonyl chloride of theformula V [Petersen, S. in: M{umlaut over (u)}ller, E. (Editor) Methodender Organischen Chemie [Methods of Organic Chemistry] (Houben-Weyl)8:102 (1952)].

After removal of the solvent, the amide derivatives of the formulae VIand X can be purified by chromatography, for example over silica gel(eluent: for example methylene chloride/methanol mixtures) and thenlyophilized.

The enzymes used for the preparation of compounds of the formulae I andII are commercially obtainable or can be obtained by known processes.The galactose transferase used in the present case for the enzymaticp(1→4)galactosylation can be obtained, for example, from Boehringer.Exclusively β-specific galactosylation of the 4-OH function of theglucosamine takes place [Paicic, M. M., Methods Enzymol. 230:300-316(1994)]. The galactose transferase used for the β(1→43)galactosylationcan be produced, for example, by genetic engineering (JPN 06181759 A2,Application JP 92-336436921216). Exclusively galactosylation on the 3-OHfunction of the N-acyl-glucosamide takes place.

The α(2→3)sialic acid transferase is preferably a microbially producedsialyl transferase (WO 91/06635), the original site of occurrence is therat liver. A strictly α-specific sialylation of the 3-OH group of theterminal galactose takes place.

The microbially produced fucose transferase VI (fuc-t VI) transfers thepyranose α-specifically to the 3-OH group of the N-acylglucosamine unit.The fucose transferase III (fuc-t III) also produced microbiallytransfers the pyranose α-specifically to the 4-OH group of theN-acylglucosamine unit (WO 91/12340).

The enzymatic reactions are advantageously carried out in the presenceof 0.1 U to 5 U of the enzyme in question. It has proved favourable toemploy the glycosyl donor in excess. Good results are achieved if, forexample, 1.2 to 2 equivalents of uridine di-phosphategalactose, 1.2 to2.3 equivalents of cytidine mono-phosphate-sialic acid or 1.2 to 2.5equivalents of guanosine di-phosphate-fucose are employed.

The UDP-galactose can be obtained commercially or synthesizedchemo-enzymatically. For this, hydroxyl-protective groups of the formula—C(O)—R of the sugar residue, in which R is linear or branched alkyl,preferably C₁-C₈alkyl, particularly preferably C₁-C₄alkyl, unsubstitutedphenyl or phenyl which is substituted by C₁-C₄alkyl or C₁-C₄alkoxy, aresplit off enzymatically from a protected UDP-galactose. Examples ofhydroxyl-protective groups are protective groups of the formula —C(O)—R,in which R is methyl, ethyl, n- or i-propyl, n-, i-, s- or t-butyl orpentyl, hexyl, heptyl or octyl with all the possible isomers, or isunsubstituted phenyl, or is phenyl which is mono- to trisubstituted inan identical or different manner with a substituent chosen from thegroup consisting of methyl, ethyl, n- and i-propyl, n-, i-, s- andt-butyl, methoxy, ethoxy, n- and i-propoxy and n-, i-, s- and t-butoxy.Examples of substituted phenyl are derived from toluene, o-, m- andp-xylene, pseudocumene, mesitylene, trimethylbenzene, ethylbenzene,dimethylpropylbenzene and cumene. This process can be carried out withsoluble or immobilized enzymes. The choice of enzyme depends on thenature of the protective groups and the stereochemistry on the sugar. Ithas proved advantageous here to use a functionally homogeneous enzyme oran enzyme mixture. If the protective group is a radical —C(O)—CH₃, it issplit off with an acetyl-esterase. If it is a radical —C(O)—CH₂CH₃, theprotective group is split off with an acetyl-esterase, a lipase or amixture of these two enzymes. Lipases are preferably employed forsplitting off the radicals —C(O)-C₃-C₈alkyl and unsubstituted orsubstituted —C(O)-phenyl. The enzymes can originate from naturallyoccurring sources, such as animals, microorganisms or plants, or alsoproduced by genetic engineering. Commercially obtainable enzymes, forexample plant enzymes, such as the acetyl-esterase from orange peel (EC3.1.1.6), are of particular advantage. The reaction can be carried outboth in the presence and in the absence of buffers. If buffers arepresent, these are advantageously electrolytic buffers, such as NaCl,MgHPO₄, 2-morpholinoethanesulfonic acid monohydrate-NaOH,N-(2-acetamino)-2-aminoethanesulfonic acid NaOH-NaCl,3-morpholinopropanesulfonic acid NaOH-NaCl,N-tris(hydroxymethyl)methyl-2-aminoethanesulfonic acid NaOH-NaCl,4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid NaOH-NaCl andimidazole HCl-NaCl. The reaction is preferably carried out in atemperature range between room temperature and 40° C., preferably at 37°C. The pH is advantageously in a range between pH 6.5 and pH 7.5, and ispreferably pH 7, and is advantageously kept constant automatically, forexample with the aid of pH probes and automatic metering equipment. Thechoice of buffer, temperature and pH otherwise depend on the particularenzyme used and the substrate to be reacted and in individual cases itis entirely possible for it to lie outside the ranges stated. Theprocess can also be carried out by activating either the sugar1-phosphate or the corresponding nucleoside with a carbonyl-bis-azolebefore the coupling and, after the coupling, splitting off theprotective groups enzymatically. Examples of carbonyl-bis-azoles arecarbonyidiimidazole, carbonylditriazole, thiocarbonyldiimidazole andcarbonyidioxydibenzotriazole. For example, protected monophosphoric acidsugar-esters are reacted with an excess of carbonyl-bis-azole in thepresence of a polar solvent. The excess carbonyldiazole is thenadvantageously destroyed with a precisely metered amount of absolutemethanol. After this activation, the activated sugar-phosphates arereacted in situ or after isolation with trialkylammonium salts of thenucleotide units to give the protected nucleoside di- ortriphosphate-sugars. The imidazole salt primarily formed is thenfiltered over an ion exchanger, to be replaced by any ion Q. Furtherpurification can then be carried out on reversed phase silica gels or byprecipitation with suitable precipitants, such as ethanol orethanol/isopropanol or ethanol/acetone mixtures. The reaction isadvantageously carried out in the absence of water in a dry, polar,non-hydroxylic solvent in a temperature range between room temperatureand 80° C., preferably in a range between 40° C. and 50° C., inparticular at 40° C. It has proved advantageous to carry out thereaction in an ultrasonic bath. Examples of polar, non-hydroxylicsolvents are dimethylformamide, dimethyl sulfoxide, acetone, dioxane,pyridine and acetonitrile and mixtures thereof.

The CMP-sialic acid donor where R₃ is methyl is commercially obtainable,but like the corresponding donor where R₃ is hydroxymethyl, canadvantageously also be prepared enzymatically [Heidlas, J. E., Williams,K. W., Whitesides, G. M., Acc. Chem. Res. 25:307-314 (1992)].

The GDP-activated donor of the formula XI for the last preparation stepcan advantageously be prepared chemo-enzymatically as described abovefor UDP-galactose.

The enzymatic transfer of galactose and sialic acid can be carried outboth in a single step and in two successive steps.

The amidations can be carried out, depending on the definition of R₁,R₂, R₄, R₇ and X, in accordance with one of the customary specifications[for example Bodansky, M., Principles of Peptide Chemistry, 2nd Edition,16-61, Springer Berlin (1993)]. For enzymatic syntheses with galactosetransferase, sialic acid transferase and fucose transferase, it isadvantageous to carry out the synthesis in the presence of buffers, suchas sodium cacodylate, tris(hydroxymethyl)aminomethane or4-(2-hydroxyethyl)-piperazine-1-ethanesulfonic acid, in each case in theoptimum pH and temperature range, for example in the range from pH 6 topH 8 and in the range from 25° C. to 37° C. It has proved particularlyadvantageous if the incubation mixture comprises salts, for example 5 to40 mM manganese(II) chloride, and auxiliary enzymes, such as alkalinephosphatase from the bovine intestine (16 to 50 U).

The compounds according to the invention have an improved bindingaffinity for the corresponding selectins. The compounds according to theinvention can be employed as antiadhesion therapeutics. In the case ofpathogenic inflammations, they can prevent the selectin receptors frombinding to activated endothelial cells on sialyl-Lewis^(a) and/orsialyl-Lewis^(x) structures on the surface of leucocytes. In the case oftissue rejections, they can block corresponding receptors of thehaematolymphoid cell system. The adhesion of metastasing cells,bacteria, viruses or other pathogens and toxins can likewise besuppressed by blocking the corresponding receptors on the cell surface.

The invention also additionally relates to the compounds according tothe invention for use in a therapeutic method for the treatment ofdiseases in warm-blooded animals, including man. The dosage onadministration to warm-blooded animals of about 70 kg body weight canbe, for example, 0.01 to 1000 mg per day. The compounds are preferablyadministered parenterally, for example intravenously orintraperitoneally, in the form of pharmaceutical preparations.

The invention furthermore relates to a pharmaceutical preparationcomprising an active amount of compound according to the invention, byitself or together with other active ingredients, a pharmaceuticalcarrier, preferably in a significant amount, and, if appropriate,adjuncts.

The pharmacologically active compounds according to the invention can beused in any form of preparations for parenteral administration orinfusion solutions. Such solutions are preferably isotonic aqueoussolutions or suspensions, it being possible for these to be preparedbefore use, for example in the case of lyophilized preparations whichcomprise the active substance by itself or together with a carrier, forexample mannitol. The pharmaceutical preparations can be sterilizedand/or comprise adjuncts, for example preservatives, stabilizers,wetting agents and/or emulsifiers, solubilizing agents, salts forregulating the osmotic pressure and/or buffers. The pharmaceuticalpreparations, which can comprise further pharmacologically activesubstances, for example antibiotics, if desired, are prepared in amanner known per se, for example by means of conventional dissolving orlyophilizing processes, and comprise about 0.1% to 90%, in particularfrom about 0.5% to about 30%, for example 1% to 5%, of activesubstance(s).

The following Examples illustrate the invention in more detail.

Abbreviations: Ac: acetyl; CMP-sia: cytidine monophosphate-sialic acid;DMF: dimethylformamide; DMSO: dimethyl sulfoxide; DPPB:1,4-bis(diphenylphosphino)butane; GDP-ara: guanosinediphosphate-α-D-arabinose; GDP-L-gal: guanosine diphosphate-L-galactose;HBPyU: O-(benzotriazol-1-yl)-N,N,N′,N′-bis(tetramethylene)uroniumhexafluorophosphate; Ph: phenyl; BSA: bovine serum albumin (Boehringer);RT: room temperature; TBTU:O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate;THF: tetrahydrofuran; UDP-gal: uridine diphosphate-D-galactose

All enzymatic steps are performed in plastic vessels.

A PREPERATION OF THE STARTING COMPOUNDS EXAMPLE A1 Preparation ofCompound No. (54)

(a) 8.63 g (20.0 mmol) ofα,β-1,3,4,6-tetra-O-acetyl-2-deoxy-2-N-allyloxycarbonyl-glucose[Boullanger, P., Jouineau, M., Bouammali, B., Lafont, D., Descotes, G.,Carbohydr. Res. 202:151-164 (1990)] are reacted by known processes[Lafont, D., Manaudier, S., Boullanger, P., Descotes, G., Bull. Soc.Chim. Fr. 127:576-583 (1990)] with 5.65 g (30.0 mmol) of methyl9-hydroxy-nonanecarboxylate [Lemieux, R. U., Bundle, D. R., Baker, D.A., J. Am. Chem. Soc. 97:4076-4083 (1975)] in the presence of 10.3 ml(56.0 mmol) of methyl trifluoromethanesulfonate at −30° C. in 150 ml ofmethylene chloride. After chromatography of the reaction mixture onsilica gel (eluent: petroleum ether/ethyl acetate—2/1), 11.14 g(quantitative) of compound No. (17) are obtained.

(b) 5.15 g (9.2 mmol) of monosaccharide No. (17) are added to 30 ml ofdry methanol, in which 15.0 mg (0.65 mmol) of sodium has been dissolvedbeforehand, at RT under an argon atmosphere. After about 1 h, the sugaris deacetylated completely. The reaction mixture is then poured onto astrongly acid ion exchanger (DOWEX 8×50 strongly acidic, Fluka), themixture is shaken for 15 minutes, the ion exchanger is filtered off andwashed again with about 100 ml of methanol and the combined organicphases are evaporated. The resulting white powder is dried under a highvacuum. 3.95 g (99%) of deprotected sugar No. (11) are obtained[({umlaut over (O)}hrlein, R., Ernst, B., Berger, E. G., Carbohydr. Res.236:335-338 (1992)].

¹H-NMR (CD₃OD, 250.13 MHz) δ=1.22 (m, 8 H); 1.47 (m, 4 H); 2.22 (t, 7.6Hz, 2 H); 3.19-3.43 (m, 5 H); 3.55 (s, 3 H); 3.60 (dd, 5.5 Hz, 10.3 Hz,1 H); 3.78 (m, 2 H); 4.25 (d, 7.3 Hz, 1 H); 4.42 (m, 2 H); 5.10 (broadd, 17.2 Hz, 1 H); 5.23 (broad d, 17.2 Hz, 1 H); 5.86 (m, 1 H). ¹³C-NMR(CD₃OD, 62.90 MHz) δ=26.00; 27.01; 30.11; 30.31; 30.34; 30.62; 34.77;51.98; 59.00; 62.79; 66.36; 70.66; 72.13; 75.93; 77.81; 103.11; 117.30;134.49; 158.88; 175.97.

(c) 9.7 g (22.4 mmol) of monosaccharide No. (11) are dissolved in 100 mlof dry THF. 6 ml (40.0 mmol) of benzaldehyde dimethylacetal and 250 mgof racemic camphor-10-sulfonic acid are added to this solution insuccession and the mixture is heated to 50° C. It is stirred overnightuntil all the starting material has been consumed and is then cooled toRT and, before the solvent is evaporated off, a further 0.5 ml oftriethylamine is added. The residue is chromatographed over silica gel(eluent: methylene chloride/methanol—20/1). 11.0 g (95%) of4,6-protected sugar No. (54) are obtained. ¹H-NMR (CDCl₃, 400.13 MHz)δ=1.20 (m, 8 H); 1.51 (m, 4 H); 2.23 (t, 7.6 Hz, 2 H); 3.25-3.50 (m, 5H); 3.60 (s, 3 H); 3.70 (t, 9.7 Hz, 1 H); 3.78 (dt, 4.8 Hz, 9.7 Hz, 1H); 4.25 (dd, 4.8 Hz, 10.9 Hz, 1 H); 4.50 (m, 2 H); 5.12 (m, 2 H); 5.23(dq, 1.2 Hz, 16.3 Hz, 1 H); 5.45 (s, 1 H); 5.84 (m, 1 H); 7.30 (m, 3 H);7.42 (m, 2 H). ¹³C-NMR (CDCl₃, 100.61 MHz) δ=24.77; 25.63; 28.91; 29.00;29.35; 34.16; 51.46; 58.60; 65.73; 66.04; 68.59; 70.21; 70.69; 72.27;81.49; 101.75; 117.60; 126.21 (2×C); 128.23 (2×C); 129.17; 132.46;159.16; 174.53.

EXAMPLE A2 Preparation of Compound No. (3)

(a) 13.1 g (82%) of amine No. (2) are obtained as a white powder from20.0 g (46 mmol) of monosaccharide No. (11) in the presence of 0.9 g oftetrakis-triphenylpalladium, 0.9 g of DPPB and 11.9 g (82.9 mmol) ofsodium thiophenolate in dioxane/methanol/THF (200 ml-40 ml-100 ml)analogously to known processes [Boullanger, P., Banoub, J., Descotes,G., Can. J. Chem. 65:1343-1348 (1987) or Gen{circumflex over (e)}t, J.P., Blart, E., Savignac, M., Lemeune, S., Lemaire-Audoire, S., Bernard,J. M., Synlett 680-682 (1993)] after chromatography of the reactionmixture over silica gel (eluent: methylene chloride/methanol—7/1).

¹H-NMR (CD₃OD, 62.90 MHz) δ=1.25 (m, 8 H); 1.51 (m, 4 H); 2.23 (t, 8.4Hz 2 H); 2.50 (bd, 8.3 Hz, 1 H); 3.19 (m, 3 H); 3.41 (dt, 4.2 Hz, 8.4Hz, 1 H); 3.57 (s, 3 H); 3.59 (bdd, 4.8 Hz, 12.4 Hz, 1 H); 3.71 (m, 2H); 4.13 (d, 7.6 Hz, 1 H).

(b) 3.35 g (9.6 mmol) of compound No. (2) are dissolved in 90 ml ofmethanol at RT, 1.09 ml of acetic anhydride and 1.60 ml of triethylamineare added in succession and the mixture is stirred overnight at RT. Thereaction mixture is evaporated and the residue is chromatographed oversilica gel (eluent: methylene chloride/methanol—7/1). 3.75 g of compoundNo. (3) are obtained as a white powder. ¹H-NMR (CD₃OD, 250.13 MHz)δ=1.22 (m, 8 H); 1.49 (m, 4 H); 1.89 (s, 3 H); 2.22 (t, 7.6 Hz, 2 H);3.18-3.86 (m, 10 H); 4.31 (d, 7.6 Hz, 1 H). ¹³C-NMR (CD₃OD, 62.98 MHz)δ=23.75; 26.70; 27.72; 30.82; 30.99 (2×C); 31.08; 31.30; 35.47; 52.69;58.05; 63.47; 71.28; 72.81; 77.78; 78.57; 103.37; 174.13.

EXAMPLE A3 Preparation of Compound No. (16)

(a) 36 mg (214 μmol) of vanillic acid are introduced into 3 ml of dryDMF, and 30 μl (216 μmol) of triethylamine and 91 mg (211 μmol) of TBTUare added at RT [Dourtoglou, V., Gross, B., Lambropoulou, V., Zioudrou,C., Synthesis 572-574 (1984)]. 100 g (286 μmol) of amine No. (2) areadded to the resulting clear solution and the mixture is stirredovernight. After the solvent has been evaporated off and the residue hasbeen chromatographed over RP-18 gel (eluent: methanol/water—1/1), 41 mg(41%) of compound No. (16) are obtained as a white powder afterlyophilization from dioxane. ¹H-NMR (CD₃OD-CDCl₃, 250.13 MHz) δ=1.10 (m,8 H); 1.46 (m, 4 H); 2.22 (t, 7.5 Hz, 2 H); 3.40-3.92 (m, 14 H); 4.59(d, 8.2 Hz, 1 H); 6.82 (d, 8.3 Hz, 1 H); 7.36 (dd, 2.1 Hz, 8.3 Hz, 1H);7.44 (d, 2.1 Hz, 1 H); ¹³C-NMR (CD₃OD-CDCl₃, 62.90 MHz) δ=25.78; 26.90;29.90; 30.13 (2×C); 30.45; 34.76; 52.14; 56.46; 57.70; 62.90; 70.67;72.11; 76.06; 77.32; 102.12; 111.96; 115.59; 121.88; 127.24; 150.14;151.49; 167.68; 170.74.

(b1) 3.9 g (76%) of free amine No. (18) are obtained in accordance withthe instructions by Boullanger et al. [Boullanger, P., Banoub, J.,Descotes, G., Can. J. Chem. 65:1343-1348 (1987)] or Gen{circumflex over(e)}t et al. [Gen{circumflex over (e)}t, J. P., Blart, E., Savignac, M.,Lemeune, S., Lemaire-Audoire, S., Bernard, J. M., Synlett 680-682(1993)] from 6.0 g (10.7 mmol) of compound No. (17).

¹H-NMR (CD₃OD, 250.13 MHz) δ=1.34 (m, 8 H); 1.64 (m, 4 H); 2.06 (s, 3H); 2.11 (s, 6 H); 2.33 (t, 7.6 Hz, 2 H); 2.95 (dd, 2.1 Hz, 8.3 Hz, 1H); 3.52 (dt, 7.6 Hz, 8.3 Hz, 1 H); 3.71 (m, 4 H); 3.93 (dt, 7.6 Hz, 8.3Hz, 1 H); 4.15 (dd, 2.1 Hz, 11.0 Hz, 1 H); 4.28 (d, 7.3 Hz, 1 H); 4.72(dd, 5.5 Hz, 11.0 Hz, 1 H); 5.02 (m, 2 H). ¹³C-NMR (CDCl₃, 62.90 MHz)δ=20.17; 20.25; 20.31; 24.37; 25.37; 28.51; 28.63 (2×C); 28.99; 33.48;50.90; 55.48; 61.82; 68.47; 69.74; 71.26; 74.90; 103.57; 169.22; 170.06;173.59.

(b2) 100 mg (210 mmol) of amine No. (18), 47 mg (280 mmol) of vanillicacid and 121 mg (280 mmol) of HBPyU are dissolved in 3 ml of absoluteacetonitrile at room temperature, 31 μl of triethylamine are then addedand the mixture is stirred for three days. After the solvent has beenevaporated off and the residue has been chromatographed over silica gel(eluent: petroleum ether/ethyl acetate—1/3), 84 mg (64%) ofperacetylated amide are obtained; the product is dissolved in 2 ml ofdry methanol at RT, and 308 μmol of freshly prepared sodium methanolateare added. After about 4 hours, the mixture is neutralized with DOWEX50×8 H⁺ form, the resin is filtered off and the mixture is evaporated.The residue is chromatographed over silica gel and the desired productis lyophilized from dioxane/water. 37 mg (65%) of amide No. (16) areobtained as a white powder.

EXAMPLE A4 Preparation of Compound No. (23)

82 mg (526 μmol) of orotic acid are suspended in 5 ml of dry DMF. 74 μlof triethylamine, 227 mg (527 μmol) of TBTU and 250 mg (716 mmol) ofamine No. (2) are added in succession under argon, with vigorousstirring. After purification by chromatography twice, first over silicagel (eluent: methylene chloride/methanol—10/2) and then over reversedphase gel (eluent: methanol/water—1/1), 123 mg (48%) of amide No. (23)are obtained as a white powder after lyophilization from dioxane/water.¹H-NMR (D₆-DMSO, 250.13 MHz) δ=1.28 (m, 8 H); 1.45 (m, 4 H); 2.26 (t,7.5 Hz, 2 H); 3.14 (m, 2 H); 3.43 (m, 4 H); 3.59 (s, 3 H); 3.70 (m, 2H); 4.39 (d, 8.2 Hz, 2 H); 6.11 (s, 1 H); 8.68 (broad d, 9.6 Hz, 1 H).¹³C-NMR (D₆-DMSO, 62.89 MHz) δ=25.12; 26.21; 29.15; 29.41; 29.48; 29.70;33.96; 51.87; 56.79; 61.56; 69.20; 71.07; 74.31; 77.71; 99.97; 101.33;147.17; 153.16; 160.98; 165.20; 174.11.

EXAMPLE A5 Preparation of Compound No. (49)

335 μl of methyl formate are added to 40 mg (114 μmol) of amine No. (2)in 1 ml of methanol and a catalytic amount of triethylamine, and themixture is heated at 50° C. for 2 days. After the mixture has beenconcentrated, the residue is chromatographed over silica gel (eluent:methylene chloride/methanol—10/2). 39 mg (91%) of formamide No. (49) areobtained as an isomer mixture (about 60/40). ¹H-NMR (CD₃OD, 250.13 MHz)δ=1.21 (m, 8 H); 1.49 (m, 4 H); 2.22 (t, 7.6 Hz, 2 H); 2.92-3.45 (m, 5H); 3.52-3.64 (m, 4 H); 3.74-3.89 (m, 2 H); 4.22 (d, 8.5 Hz, 0.4 H);4.35 (d, 8.5 Hz, 0.6 H); 7.85 (s, 0.4 H); 8.05 (s, 0.6 H). M: mainisomer; S: secondary isomer; ¹³C-NMR (CD₃OD, 62.90 MHz) δ=25.99; 26.99;30.09; 30.26; 30.33; 30.58; 34.76; 51.98; 56.31 M; 60.71 S; 62.59 S;62.71 M; 70.59 M; 70.83 S; 71.89 S; 72.02 M; 75.38 S; 75.80 M; 77.76 S;77.91 M; 102.30 S; 102.42 M; 164.12 M; 168.14 S; 176.02.

EXAMPLE A6 Preparation of Compound No. (78)

1.25 g (36 mmol) of amine No. (2) are dissolved in 45 ml of absolutemethylene chloride at RT, and 310 μl (33 mmol) of ethyl chloroformateand 45 μl of triethylamine are added in succession. After about 5 hours,the reaction mixture is evaporated and the residue is chromatographedover silica gel (eluent: methylene chloride/methanol—15/2). 950 mg (69%)of monosaccharide No. (78) are obtained. ¹H-NMR (CD₃OD, 250.13 MHz)δ=1.18 (t, 7.5 Hz, 3 H); 1.25 (m, 8 H); 1.49 (m, 4 H); 2.23(t, 7.6 Hz, 2H); 3.10-3.46 (m, 5 H); 3.57 (s, 3 H); 3.60 (dd, 5.5 Hz, 10.0 Hz, 1 H);3.74-3.88 (m, 2 H); 4.00 (q, 7.5 Hz, 2 H); 4.26 (broad d, 8.6 Hz, 1 H).¹³C-NMR (CD₃OD, 62.9 Mhz) DEPT δ=14.37; 25.33; 26.31; 29.43; 29.62;29.67; 29.93; 34.10; 51.35; 58.21; 61.06; 62.08; 70.02; 71.42; 75.25;77.11; 102.48.

EXAMPLE A7 Preparation of Compound No. (95)

(a) 1.3 g (3.7 mmol) of amine No. (2) are dissolved in 20 ml of methanolat RT, and 1.0 ml (6.1 mmol) of triethylamine and 0.8 ml (5.2 mmol) ofbenzyloxyacetyl chloride are added in succession and the mixture isstirred overnight. The solvent is now evaporated off and the residue ischromatographed over silica gel (eluent: methylenechloride/methanol—9/1). 1.6 g (85%) of amide No. (94) are obtained.

¹H-NMR (CD₃OD, 250.13 MHz) δ=1.21 (m, 8 H); 1.48 (m, 4 H); 2.21 (t, 7.5Hz, 2 H); 3.23 (m, 2 H); 3.39 (dt, 6.9 Hz, 9.0 Hz, 1 H); 3.47-3.70 (m, 6H); 3.81 (m, 2 H); 3.91 (m, 2 H); 4.44 (d, 8.6 Hz, 1 H); 4.56 (s, 2 H);7.30 (m, 5 H). ¹³C-NMR (CD₃OD, 62.9 MHz) δ=25.97; 27.02; 30.09; 30.28;30.31; 30.62; 34.75; 51.97; 57.11; 62.79; 70.23; 70.52; 72.22; 74.22;75.61; 77.87; 102.36; 129.09; 129.15; 129.53; 136.64; 172.81; 175.89.

(b) 1.6 g (3.2 mmol) of compound No. (94) are dissolved in 50 ml ofmethanol at RT to give a clear solution, and 200 mg of 10%palladium-on-charcoal are added under an argon atmosphere. Hydrogen ispassed through this mixture, with vigorous stirring, until no furtherdeduct is detectable. The charcoal is now filtered off over Celite, thesolvent is evaporated and the residue which remains is dried under ahigh vacuum. 1.2 g (91%) of monosaccharide No. (95) are obtained as acolourless solid. ¹H-NMR (CD₃OD, 250.13 MHz) δ=1.25 (m, 8 H); 1.49 (m, 4H); 2.22 (t, 7.5 Hz, 2 H); 3.16-3.45 (m, 9 H); 3.71-3.81 (m, 2 H); 3.92(m, 2 H); 4.40 (d, 8.6 Hz, 1 H). ¹³C-NMR (CD₃OD, 62.9 MHz) δ=26.00;26.97; 30.10; 30.29 (2×C); 30.61; 34.78; 51.96; 57.10; 62.73; 62.81;70.56; 72.22; 75.78; 77.87; 102.45; 175.55; 176.05.

EXAMPLE A8 Preparation of Compound No. (28)

65 mg (64%) of monosaccharide No. (28) are obtained from 31 mg (204μmol) of 2-hydroxy-5-methylbenzoic acid and 100 mg (210 μmol) of amineNo. (18) in the presence of 95 mg (220 μmol) of HBPyU in 3 ml of dryacetonitrile. ¹H-NMR (CD₃OD, 250.13 MHz) δ=1.03 m, 8 H); 1.39 (m, 4 H);2.15 (t, 7.6 Hz, 2 H); 2.21 (s, 3 H); 3.23-3.44 (m, 3 H); 3.57 (s, 3 H);3.62 (m, 2 H); 3.81 (m, 3 H); 4.50 (d, 7.6 Hz, 1 H); 6.71 (d, 7.6 Hz, 1H); 7.10 (dd, 1.4 Hz, 7.6 Hz, 1 H); 7.53 (d, 1.4 Hz, 1 H). ¹³C-NMR(CD₃OD, 62.90 MHz) δ=20.62; 25.79; 26.90; 29.86; 30.03; 30.09; 30.38;34.68; 51.94; 57.15; 62.64; 70.60; 72.07; 75.42; 77.60; 102.54; 116.22;118.21; 128.34; 128.91; 135.41; 159.18; 171.62; 175.91.

EXAMPLE A9 Preparation of Compound No. (33)

86 mg (84%) of compound No. (33) are obtained according to Example A8from 37 mg (240 μmol) of 3,5-dihydroxybenzoic acid and 100 mg (210 μmol)of compound No. (18) in the presence of 104 mg (240 μmol) of HBPyU.¹H-NMR (CD₃OD, 250.13 MHz) δ=1.17 (m, 8 H); 1.49 (m, 4 H); 2.16 (t, 7.5Hz, 2 H); 3.34-3.55 (m, 3 H); 3.59-3.96 (m, 8 H); 4.55 (d, 8.6 Hz, 1 H);6.43 (t, about 2.0 Hz, 1 H); 6.76 (d, about 2.0 Hz, 2 H). ¹³C-NMR(CD₃OD, 62.90 MHz) δ=25.97; 27.15; 30.07; 30.26; 30.30; 30.54; 34.78;51.94; 57.92; 62.54; 70.70; 72.31; 75.70; 77.92; 102.78; 106.64; 107.19(2×C); 138.15; 159.72 (2×C); 169.66; 176.22.

EXAMPLE A10 Preparation of Compound No. (37a)

47 mg (47%) of monosaccharide No. (37a) are obtained according toExample A8 from 34 mg (220 μmol) of 3-fluoro-6-hydroxybenzoic acid and100 mg (210 μmol) of amine No. (18). ¹H-NMR (CD₃OD-CDCl₃, 250.13 MHz)δ=1.09 (m, 8 H); 1.45 (m, 4 H); 2.21 (t, 7.6 Hz, 2 H); 3.33-3.89 (m, 11H); 4.54 (d, 7.6 Hz, 1 H); 6.84 (dd, 5.5 Hz, 10.3 Hz, 1 H); 7.02 (ddd,3.4 Hz, 7.6 Hz, 8.3 Hz, 1 H); 7.42 (dd, 5.5 Hz, 10.3 Hz, 1 H). ¹³C-NMR(CD₃OD-CDCl₃, 62.90 MHz) δ=24.61; 25.57; 28.72; 28.82; 28.85; 29.15;33.84; 51.37; 55.84; 61.26; 70.07; 70.58; 74.03; 75.27; 100.94; 112.97(d, 24.2 Hz); 115.24 (d, 6.5 Hz); 118.69; 120.90 (d, 23.4 Hz); 155.11(d, 174.2 Hz); 169.40; 174.80. ¹⁹F-NMR (CD₃OD-CDCl₃, 235.36 MHz)δ=−73.36.

EXAMPLE A11 Preparation of Compound No. (41)

47 mg (47%) of monosaccharide No. (41) are obtained according to ExampleA8 from 42 mg (220 μmol) of 8-hydroxy-quinoline-2-carboxylic acid and100 mg (210 μmol) of amine No. (18) after deacetylation with 1.05equivalents of sodium methanolate. ¹H-NMR (CD₃OD, 250.13 MHz)δ=0.38-1.40 (m, 12 H); 1.92 (t, 7.6 Hz, 2 H); 3.28-3.92 (m, 11 H); 4.48(d, 7.6 Hz, 1 H); 7.09 (dd, 0.9 Hz, 7.6 Hz, 1 H); 7.34 (dd, 0.9 Hz, 7.6Hz, 1 H); 7.47 (t, 7.6 Hz, 1 H); 8.12 (d, 8.3 Hz, 1 H); 8.31 (d, 8.3 Hz,1 H). ¹³C-NMR (CD₃OD, 62.90 MHz) δ=26.52; 27.09; 29.89; 30.17; 30.22;30.50; 34.64; 51.92; 57.81; 62.82; 70.61; 72.23; 75.94; 78.05; 102.88;112.78; 118.99; 120.15; 130.57; 131.46; 138.34; 138.82; 148.89; 155.02;167.04; 175.87.

EXAMPLE A12 Preparation of Compound No. (45)

97 mg (82%) of monosaccharide No. (45) are obtained according to ExampleA8 from 44 mg (240 μmol) of 3-hydroxy-4-nitrobenzoic acid and 110 mg(231 μmol) of amine No. (18) in the presence of 100 mg of HBPyU and 31μl of triethylamine. ¹H-NMR (CD₃OD-CDCl₃, 250.13 MHz) δ=1.22 (m, 8 H);1.59 (m, 4 H); 2.35 (t, 7.6 Hz, 2 H); 3.39-3.65 (m, 3 H); 3.71-4.07 (m,8 H); 4.70 (d, 7.6 Hz, 1 H); 7.53 (dd, 2.1 Hz, 8.3 Hz, 1 H); 7.70 (d,2.1 Hz, 1 H); 8.23 (d, 8.3 Hz, 1 H). ¹³C-NMR (CD₃OD-CDCl₃, 62.90 MHz)δ=25.55; 29.67; 29.86 (2×C); 30.12; 34.69; 52.30; 57.41; 61.90; 70.84;71.19; 74.59; 76.93; 102.02; 119.48; 119.77; 126.29; 136.53; 143.03;154.68; 167.82; 176.04.

PREPERATION OF THE MIMETICS EXAMPLE B1.1 Preparation of Compound No.(10)

(a1) Galactosylation with β(1→4)galactose transferase 512.0 mg (24.7μmol) of compound No. (11), 31.5 μmol of UDP-gal, 1.4 mg of BSA and 16.9mg (85 μmol) of manganese(II) chloride hexahydrate are brought togetherin 1.0 ml of sodium cacodylate buffer (0.1 M, pH=7.43) and the mixtureis briefly treated with ultrasound in an ultrasonic bath. 0.2 U ofgalactose transferase (Sigma; 0.2 ml) and 34 U (2 μl) of alkalinephosphatase from the bovine intestine (Boehringer) are added to theresulting homogeneous, milky suspension. The mixture is mixed andincubated at 37° C., while stirring. The reaction precipitates arecentrifuged off, the clear supernatant is lyophilized from water/dioxaneand the residue is purified by chromatography over silica gel (eluent:methylene chloride/methanol/water mixtures). The solvent is removed, theresidue is taken up in dioxane/water and renewed lyophilization gives17.5 mg of compound No. (12) (100%) as a white powder.

(a2) Galactosylation with β(1→4)galactose transferase and UDP-galactoseepimerase 1.18 mmol of compound No. (11), 1.40 mmol of uridinediphosphate-glucose (UDP-glc) (Sigma), 8.9 mg of BSA and 64.0 mg (323μmol) of manganese(II) chloride hexahydrate (Fluka) are brought togetherin 8 ml of sodium cacodylate buffer (0.1 M, pH=7.52) and the mixture isbriefly treated with ultrasound in an ultrasonic bath. 24 U ofgalactosyl transferase (6 ml), 800 μl of UDP-galactose epimerase (Sigma,100 U/2 ml) and 129 U (8 μl) of alkaline phosphatase from the bovineintestine (Boehringer) are added to the resulting homogeneous, milkysuspension. The mixture is mixed and incubated at 37° C., whilestirring. At the end of the reaction, the reaction precipitates arecentrifuged off, the clear supernatant is lyophilized from water/dioxaneand the residue is purified by chromatography over silica gel (eluent:methylene chloride/methanol/water mixtures). The solvent is removed, theresidue is taken up in dioxane/water and renewed lyophilization gives621 mg (88%) of compound No. (12) as a white powder. ¹H-NMR(CD₃OD-CDCl₃, 250.13 MHz) δ=1.22 (m, 8 H); 1.48 (m, 4 H); 2.21 (t, 7.5Hz, 2 H); 3.24-3.85 (m, 17 H); 4.29 (broad d, 8.6 Hz, 2 H); 4.44 (m,2H); 5.08 (dd, 10.3 Hz, 1.4 Hz, 1 H); 5.23 (broad d, 15.1 Hz, 1 H); 5.82(m, 1 H). ¹³C-NMR (CD₃OD-CDCl₃, 62.89 MHz) δ=25.85; 26.83; 29.97; 30.15;30.19; 30.45; 34.69; 51.96; 58.22; 61.84; 62.38; 66.25; 70.11; 70.65;72.41; 73.99; 74.58; 76.17; 76.89; 80.91; 102.91; 104.85; 117.28;134.26; 158.52; 175.86.

(b) Sialidation with α(2→3) sialic acid transferase 202 mg (339 μmol) ofcompound No. (12) are added to a mixture of 3 ml of a manganeses(II)chloride solution (0.06 M), 3 ml of sodium cacodylate buffer (0.05 M,pH=6.5) and 2.0 ml of doubly distilled water in a plastic test tube. Themixture is briefly treated with ultrasound in an ultrasonic bath. 329 mg(499 μmol) of CMP-sia (content about 90%), 3.5 mg of BSA, 300 μl (2.1 U)of sialyl-transferase and 4 μl (64 U) of alkaline phosphatase from thebovine intestine (Boehringer) are then added, the components are mixedand the mixture is incubated at 37° C., while stirring. At the end ofthe reaction, the reaction precipitates are centrifuged off. The clearsupernatant is filtered over a reversed phase C-18 column (eluent:methanol) and then purified over a silica gel column (eluent: methylenechloride/methanol/water mixtures). The solvent is removed, the residueis taken up in dioxane/water and the product is lyophilized. 186 mg ofcompound No. (13) (72%) are obtained as a white powder.

¹H-NMR (CD₃OD, 250.13 MHz) δ=1.20 (m, 8 H); 1.44 (m, 4 H); 1.68 (broadt, 11. 6 Hz, 1 H); 1.92 (s, 3 H); 2.19 (t, 7.6 Hz, 2 H); 2.70 (dd, 11.6Hz, 2.8 Hz, 1 H); 3.21-3.88 (m, 23 H); 3.93 (dd, 10.3 Hz, 2.1 Hz, 1 H);4.25 (broad d, 8.3 Hz, 1 H); 4.35 (broad d, 8.4 Hz, 1 H); 4.42 (m, 2 H);5.06 (m, 1 H); 5.20 (m, 1 H); 5.81 (m, 1 H). ¹³C-NMR (CD₃OD, 62.98 MHz)δ=22.66; 26.01; 26.99; 30.12; 30.29; 30.33; 30.62; 34.79; 41.95; 51.96;53.97; 58.44; 62.04; 62.73; 64.42; 66.37; 69.11; 69.28; 70.03; 70.78;70.85; 72.96; 74.27; 74.92; 76.42; 77.02; 77.61; 81.30; 101.13; 103.18;104.99; 117.27; 134.51; 158.83; 175.05; 175.49; 176.53.

(c) Fucosylation with fucose transferase VI 23.3 mg (26.2 μmol) ofcompound No. (13), 25.0 mg (38.5 μmol) of GDP-gal and 1.3 mg of BSA areadded to a mixture of 150 μl of manganese(II) chloride solution (0.25M), 450 μl of sodium cacodylate buffer (0.25 M, pH=6.48) and 600 μl ofdouble-distilled water. 2 μl (32 U) of alkaline phosphatase from thebovine intestine (Boehringer) and 250 μl (500 mU) of a fucosetransferase VI solution are added, the components are mixed and themixture is incubated at 37° C., while stirring. At the end of thereaction, the reaction precipitates are centrifuged off and the clearsupernatant is passed over a reversed phase C-18 column (eluent:methanol). The product-containing fractions are lyophilized fromwater/dioxane, filtered over an Na⁺ column (Dowex) and lyophilizedagain. Finally, the residue is purified over a silica gel column(eluent: methylene chloride/methanol/water mixtures) and lyophilizedagain from water/dioxane. 18 mg of compound No. (10) (66%) are obtainedas a white powder. ¹H-NMR (CD₃OD, 250.13 MHz) δ=1.24 (m, 8 H); 1.49 (m,4 H); 1.66 (broad t, 12.4 Hz, 1 H); 1.96 (s, 3 H); 2.24 (t, 8.4 Hz, 2H); 2.78 (dd, 12.4 Hz, 3.4 Hz, 1 H); 3.27-3.93 (m, 29 H); 4.00 (dd, 10.3Hz, 2.1 Hz, 1 H); 4.31 (d, 8.3 Hz, 1 H); 4.48 (m, 3 H); 4.71 (t, 6.2 Hz,1 H); 5.11 (m, 1 H); 5.23 (m, 1 H); 5.88 (m, 1 H). ¹³C-NMR (CD₃OD,100.62 MHz) δ=22.6; 26.0; 27.0; 30.1; 30.3; 30.4; 30.6; 34.8; 42.2;52.0; 54.0; 59.3; 61.1; 62.3; 62.6; 64.6; 66.6; 69.0; 69.3; 70.1; 70.3;70.8 (3×C); 70.9; 71.2; 73.1; 75.0; 75.7; 76.7; 77.1; 77.2; 77.7; 100.4;100.9; 102.8; 104.0; 117.4; 134.6; 158.8; 174.9; 175.5; 176.1.

EXAMPLE B1.2 Preparation of Compound No. (14)

8.0 mg (66%) of compound No. (14) are obtained from 10.7 mg (12 μmol) ofcompound No. (13) and 10.5 mg (17 μmol) of guanosinediphosphate-D-arabinose analogously to Example 6(c). ¹H-NMR (CD₃OD,250.13 MHz) δ=1.29 (m, 8 H); 1.56 (m, 4 H); 1.69 (broad t, 12.4 Hz, 1H); 1.96 (s, 3 H); 2.30 (t, 8.4 Hz, 2 H); 2.75 (dd, 12.4 Hz, 3.4 Hz, 1H); 3.46-34.05 (m, 29 H); 4.19 (dd, 8.2 Hz, 3.4 Hz, 1 H); 4.46-4.72 (m,4 H); 5.27 (dd, 8.9 Hz, 1.4 Hz, 1 H); 5.35 (dd, 17.0 Hz, 1.5 Hz, 1 H);5.96 (m, 1 H). ¹³C-NMR (CD₃OD, 62.98 MHz) δ=24.15; 26.50; 27.19; 30.34(2×C); 30.48; 31.37; 35.92; 42.07; 54.12; 54.34; 60.06; 63.42; 65.00;65.06; 65.83; 69.47; 70.08; 70.53 (2×C); 71.20; 72.94; 72.99; 73.67;75.60 (2×C); 76.24; 76.69; 76.81 (2×C); 77.14; 77.82; 100.84; 101.92;103.64; 103.87; 119.89; 134.62; 159.89; 176.19; 175.5; 176.1.

EXAMPLE B2.1 Preparation of Compound No. (1)

(a) Galactosylation with β(1→3)galactose transferase 6.8 mg (17.4 μmol)of compound No. (3), dissolved in 35 μl of DMSO, 13.8 mg (12.7 μmol) ofUDP-gal, 0.9 mg of BSA and 28 μl of a 0.5 M manganese(II) chloridesolution are added to 25 μl of sodium cacodylate buffer (0.05 M,pH=6.45). 600 μl (0.4 U/0.5 ml) of galactose transferase (JP 92-336436921216) and 33 U (2 μl) of alkaline phosphatase from the bovineintestine (Boehringer) are added to the resulting homogeneous, milkysuspension. The mixture is mixed briefly and incubated at 37° C., whilestirring. At the end of the reaction, the reaction precipitates arecentrifuged off, the clear supernatant is lyophilized from water/dioxaneand the residue is purified by chromatography over silica gel (eluent:methylene chloride/methanol/water mixtures). The solvent is removed, theresidue is taken up in dioxane/water and renewed lyophilization gives9.3 mg (97%) of compound No. (4).

¹H-NMR (CD₃OD, 250.13 MHz) δ=1.28 (m, 8 H); 1.49 (m, 4 H); 1.91 (s, 3H); 2.27 (t, 7.6 Hz, 2 H); 3.30-3.88 (m, 17 H); 4.21 (d, 7.6 Hz, 1 H);4.42 (d, 8.6 Hz, 1 H). ¹³C-NMR (CD₃OD, 62.98 MHz) δ=23.17; 26.02; 27.03;30.13; 30.28; 30.38; 30.59; 34.78; 52.55; 56.35; 62.50; 62.72; 70.21;70.59 (2×C); 72.37; 74.65; 77.13; 77.57; 84.98; 102.38; 105.59; 174.11;176.01.

(b) Sialidation with α(2→3)sialic acid transferase 81.2 mg (70%) ofcompound No. (5) are obtained as a white powder according to ExampleB1.1(b) from 76 mg (13.7 μmol) of compound No. (4).

¹H-NMR (CD₃OD, 250.13 MHz) δ=1.23 (m, 8 H); 1.49 (m, 4 H); 1.68 (broadt, 11.0 Hz, 1 H); 1.89 (s, 3 H); 1.93 (s, 3 H); 2.12 (t, 7.6 Hz, 2 H);2.73 (dd, 11.0 Hz, 4.5 Hz, 1 H); 3.28-3.90 (m, 24 H); 3.96 (dd, 8.3 Hz,3.4 Hz, 1 H); 4.27 (d, 7.6 Hz, 1 H); 4.39 (d, 7.6 Hz, 1 H). ¹³C-NMR(CD₃OD, 62.98 MHz) δ=22.74; 23.44; 26.00; 27.01; 30.12; 30.28; 30.38;30.57; 34.77; 41.79; 51.99; 53.98; 56.08; 62.69 (2×C); 64.24; 69.20;69.88; 70.50 (2×C); 72.92; 74.88; 76.75; 77.52 (3×C); 85.09; 101.17;102.40; 105.49; 174.18; 175.50; 175.99 (2×C).

(c) Fucosylation with fucose transferase III 10.7 mg (12.7 μmol) ofcompound No. (5), 13.7 mg (22.1 μmol) of GDP-ara and 1.7 mg of BSA areadded to a mixture of 150 μl of manganese(II) chloride solution (0.25M), 450 μl of sodium cacodylate buffer (0.25 M, pH=6.48) and 600 μl ofdoubly distilled water. 2 μl (32 U) of alkaline phosphatase from thebovine intestine (Boehringer) and 166 μl (100 mU) of a fucosetransferase III solution are added, the components are mixed and themixture is incubated at 37° C., while stirring. At the end of thereaction, the reaction precipitates are centrifuged off and the clearsupernatant is passed over a reversed phase C-18 column (eluent:methanol). The product-containing fractions are lyophilized fromwater/dioxane, filtered over an Na⁺ column (Dowex) and lyophilizedagain. Finally, the residue is purified over a silica gel column(eluent: methylene chloride/methanol/water mixtures) and the product islyophilized again from water/dioxane. Compound No. (1) is obtained as awhite powder (9.2 mg) (75%). ¹H-NMR (CD₃OD, 250.13 MHz) δ=1.27 (m, 8 H);1.51 (m, 4 H); 1.63 (broad t, 11.0 Hz, 1 H); 1.91 (s, 3 H); 1.93 (s, 3H); 2.25 (t, 7.6 Hz, 2 H); 2.80 (dd, 11.0 Hz, 4.5 Hz, 1 H); 3.26-4.03(m, 28 H); 4.38 (d, 8.6 Hz, 1 H); 4.41 (d, 8.6 Hz, 1 H); 4.75 (broad d,11.6 Hz, 1 H); 5.01 (d, 3.4 Hz, 1 H). ¹³C-NMR (CD₃OD, 62.98 MHz)δ=22.99; 23.50; 26.01; 27.03; 30.12; 30.27; 30.36; 30.60; 34.79; 42.50;51.98; 53.91; 57.38; 61.41; 62.77; 64.43; 65.35; 68.39; 69.37; 69.97;70.32 (2×C); 70.64 (2×C); 71.07; 72.81; 74.19; 74.86; 76.76; 77.38;77.81 (2×C); 100.29; 100.93; 102.31; 104.27; 174.00; 174.73; 175.44;176.05.

EXAMPLE B2.2 Preparation of Compound No. (6)

12.0 mg (88%) of compound No. (6) are obtained from 11.4 mg (14 μmol) ofcompound No. (5) and 14.8 mg (23 μmol) of GDP-2-fluoro-fucoseanalogously to Example B2.1. ¹H-NMR (CD₃OD, 250.13 MHz) δ=1.11 (d, 7.5Hz, 3 H); 1.26 (m, 8 H); 1.48 (m, 4 H); 1.64 (broad t, 11.0 Hz, 1 H);1.93 (s, 3 H); 1.95 (s, 3 H); 2.22 (t, 7.6 Hz, 2 H); 2.80 (dd, 11.0 Hz,4.5 Hz, 1 H); 3.25-4.08 (m, 26 H); 4.39 (d, 8.6 Hz, 1 H); 4.41 (d, 8.6Hz, 1 H); 4.51 (m, 1 H); 4.85 (broad q, 7.5 Hz, 1 H); 5.19 (d, 3.4 Hz, 1H). ¹³C-NMR (CD₃OD, 62.98 MHz) δ=16.37; 22.61; 23.56; 26.02; 27.02;30.13; 30.27; 30.37; 30.60; 34.79; 41.83; 51.97; 53.88; 57.51; 60.97;63.12; 64.45; 67.85; 68.30; 69.30 (d); 69.48; 70.00; 70.65; 70.86;72.85; 74.08; 74.30 (d); 74.81; 76.60; 77.45; 77.89; 77.95; 90.57 (d);97.44 (d); 101.42; 102.42; 104.74; 173.98; 174.92; 175.26; 176.96.

EXAMPLE B2.3 Preparation of Compound No. (7)

7.6 mg (48%) of compound No. (7) are obtained from 13.4 mg (16 μmol) ofcompound No. (5) and 13.1 mg (21 μmol) of GDP-2-amino-fucose analogouslyto Example B2.1. ¹H-NMR (CD₃OD, 250.13 MHz) δ=1.12 (d, 7.5 Hz, 3 H);1.21 (m, 8 H); 1.49 (m, 4 H); 1.63 (broad t, 11.0 Hz, 1 H); 1.91 (s, 3H); 1.93 (s, 3 H); 2.23 (t, 7.6 Hz, 2 H); 2.78 (dd, 11.0 Hz, 1 H); 3.14(dd, 10.3 Hz, 5.5 Hz, 1 H); 3.29-3.86 (m, 23 H); 3.91 (m, 2 H); 4.13 (t,8.3 Hz, 1 H); 4.40 (d, 8.6 Hz, 1 H); 4.50 (d, 8.6 Hz, 1 H); 4.61 (broadq, 7.5 Hz, 1 H); 5.16 (d, 3.4 Hz, 1 H). ¹³C-NMR (CD₃OD, 62.98 MHz)δ=16.58; 22.59; 23.56; 26.02; 27.03; 30.14; 30.28; 30.38; 30.61; 34.79;42.32; 51.97; 52.52; 53.92; 58.19; 63.05; 64.58; 68.42; 68.66; 69.15;69.43; 70.09; 70.59; 70.80; 71.98; 72.56; 72.88; 73.83; 74.86; 76.60;76.68; 76.89; 77.88; 100.94 (2×C); 101.82; 104.38; 173.99; 174.96;175.41; 176.01.

EXAMPLE B2.4 Preparation of Compound No. (8)

9.9 mg (83%) of compound No. (8) are obtained from 10.1 mg (12 μmol) ofcompound No. (5) and 14.0 mg (22 μmol) of GDP-L-galactose analogously toExample B2.1. ¹H-NMR (CD₃OD, 250.13 MHz) δ=1.27 (m, 8 H); 1.52 (m, 4 H);1.65 (broad t, 11.0 Hz, 1 H); 1.95 (s, 3 H); 1.97 (s, 3 H); 2.26 (t, 7.6Hz, 2 H); 2.81 (dd, 11.0 Hz, 4.5 Hz, 1 H); 3.29-3.99 (m, 29 H); 4.39 (d,7.6 Hz, 1 H); 4.42 (d, 7.6 Hz, 1 H); 4.72 (t, 7.5 Hz, 1 H); 5.03 (d, 3.4Hz, 1 H). ¹³C-NMR (CD₃OD, 62.98 MHz) δ=22.61; 23.62; 26.01; 27.03;30.12; 30.28; 30.36; 30.61; 34.79; 42.51; 51.97; 53.91; 57.46; 61.38;62.40; 62.81; 64.48; 68.51; 69.42; 70.02; 70.33 (2×C); 70.63 (2×C);70.95; 71.14; 72.76; 74.28; 74.85; 76.70; 77.39; 77.71; 78.65; 99.93;101.02; 102.21; 104.81; 174.10; 174.75; 175.43; 176.04.

EXAMPLE B2.5 Preparation of Compound No. (9)

7.1 mg (52%) of compound No. (9) are obtained from 11.5 mg (14 μmol) ofcompound No. (5) and 11.2 mg (17 lrmol) of GDP-L-glucose analogously toExample B2.1. ¹H-NMR (CD₃OD, 250.13 MHz) δ=1.29 (m, 8 H); 1.53 (m, 4 H);1.66 (broad t, 11.0 Hz, 1 H); 1.97 (s, 3 H); 1.99 (s, 3 H); 2.29 (t, 7.6Hz, 2 H); 2.72 (dd, 11.0 Hz, 4.5 Hz, 1 H); 3.16-3.98 (m, 30 H); 4.40 (d,7.6 Hz, 1 H); 4.46 (d, 7.6 Hz, 1 H); 5.06 (d, 3.4 Hz, 1 H). ¹³C-NMR(CD₃OD, 62.98 MHz) δ=22.60; 23.52; 26.02; 27.03; 30.13; 30.27; 30.37;30.60; 34.79; 42.33; 51.97; 53.89; 57.34; 61.32; 62.16; 62.84; 64.44;68.84; 69.46; 69.99; 70.64; 70.79; 72.17; 72.73 (2×C); 72.94; 73.73;74.55; 74.82; 76.97; 77.43; 77.72; 78.45; 98.92; 101.09; 102.46; 105.14;174.02; 174.86; 175.38; 176.55.

EXAMPLE B3.1 Preparation of Compound No. (15)

(a) 27 mg (100%) of disaccharide No. (19) are obtained according toExample B1.1(a1) from 21 mg (33 μmol) of compound No. (16) and 32 mg (52μmol) of UDP-gal (in this case the incubation mixture comprises 12% ofDMSO (vol/vol)).

¹H-NMR (CD₃OD-CDCl₃, 250.13 MHz) δ=1.05 (m, 8 H); 1.40 (m, 4 H); 2.17(t, 7.5 Hz, 2 H); 3.35-3.92 (m, 20 H); 4.35 (d, 8.3 Hz, 1 H); 4.57 (d,8.2 Hz, 1 H); 6.79 (d, 8.3 Hz, 1 H); 7.31 (dd, 2.1 Hz, 8.3 Hz, 1 H);7.39 (d, 2.1 Hz, 1 H); ¹³C-NMR (CD₃OD-CDCl₃, 100.61 MHz) δ=25.60; 26.63;29.71; 29.89; 29.95; 34.72; 52.32; 56.46; 56.81; 61.23; 61.82; 69.55;70.68; 71.95; 73.00; 73.83; 75.71; 76.38; 80.05; 102.70; 104.13; 111.48;114.93; 121.38; 126.40; 146.93; 150.11; 168.85; 175.98.

(b) 32 mg (86%) of compound No. (20) are obtained according to ExampleB1.1(b) (in this case the buffer solution comprises 9% of DMSO(vol/vol)) from 26 mg (39 μmol) of compound No. (19) and 39 mg (59 μmol)of CMP-sia.

¹H-NMR (CD₃OD, 250.13 MHz) δ=1.02 (m, 8 H); 1.48 (m, 4 H); 1.68 (broadt, 11.6 Hz, 1 H); 1.94 (s, 3 H); 2.14 (t, 7.6 Hz, 2 H); 2.76 (dd, 11.6Hz, 2.8 Hz, 1 H); 3.32-4.01 (m, 27 H); 4.38 (d, 8.6 Hz, 1 H); 4.48 (d,8.6 Hz, 1 H); 6.73 (d, 8.3 Hz, 1 H); 7.30 (dd, 2.1 Hz, 8.3 Hz, 1H); 7.39(d, 2.1 Hz, 1 H); ¹³C-NMR (CD₃OD-CDCl₃, 62.90 MHz) δ=22.70; 25.91;27.07; 30.01; 30.27 (2×C); 30.58; 34.72; 42.06; 51.90; 53.89; 56.44;57.03; 62.03; 62.66; 64.05; 69.15; 69.92; 72.91; 74.03; 74.84; 76.42;76.89; 77.50; 78.67; 79.20; 79.72; 81.77; 101.05; 102.90; 104.97;112.11; 115.72; 122.10; 127.03; 148.66; 151.18; 170.12; 175.04; 175.45;176.05.

(c) 10 mg (81%) of compound No. (15) are obtained according to ExampleB1.1(c) from 11 mg (11 μmol) of compound No. (20) and 11 mg (18 μmol) ofGDP-arabinose. ¹H-NMR (CD₃OD, 400.13 MHz) δ=0.96-1.58 (very broad m, 12H); 1.68 (broad t, 11.0 Hz, 1 (s, 3 H); 2.20 (t, 7.6 Hz, 2 H); 2.84 (dd,11.6 Hz, 2.8 Hz, 1 H); 3.36 (dd, 3.0 Hz, 10.8 Hz, 1 H); 3.39-4.11 (m, 27H); 4.51 (d, 8.6 Hz, 1 H); 4.60 (broad d, 8.6 Hz, 1 H); 5.10 (d, 3.6 Hz,1 H); 6.78 (d, 8.3 Hz, 1 H); 7.33 (dd, 2.1 Hz, 8.3 Hz,1H); 7.42 (d, 2.1Hz, 1 H); ¹³C-NMR (CD₃OD, 100.60 MHz) δ=22.57; 25.97; 27.17; 30.07;30.31; 30.33; 30.66; 34.76; 42.37; 51.95; 53.97; 56.47; 57.96; 61.40;62.96; 64.67; 65.15; 68.84; 69.31; 70.14; 70.21 (2×C); 70.72; 70.91;70.99; 73.03; 75.01; 75.44; 75.90; 76.80; 77.31; 77.88; 99.89; 100.86;102.62; 103.81; 112.28; 115.91; 122.20; 126.75; 148.91; 151.72; 170.29;174.82; 175.50; 176.09.

EXAMPLE B3.2 Preparation of Compound No. (21)

8 mg (68%) of compound No. (21) are obtained according to ExampleB3.1(c) from 10 mg (10 μmol) of compound No. (20) and 12 mg (17 μmol) ofGDP-L-galactose. ¹H-NMR (CD₃OD, 250.13 MHz) δ=0.96 (m, 8 H); 1.39 (m, 4H); 1.42 (broad t, 11.0 Hz, 1 H); 1.92 (s, 3 H); 2.16 (t, 7.6 Hz, 2 H);2.80 (dd, 11.6 Hz, 2.8 Hz, 1 H); 3.31-4.10 (m, 32 H); 4.49 (d, 8.6 Hz, 1H); 4.53 (broad d, 8.6 Hz, 1 H); 4.65 (broad t, 6.9 Hz, 1 H); 5.04 (d,5.5 Hz, 1 H); 6.74 (d, 8.3 Hz, 1 H); 7.29 (dd, 2.1 Hz, 8.3 Hz, 1 H);7.39 (d, 2.1 Hz, 1 H); ¹³C-NMR (CD₃OD, 62.90 MHz) δ=22.57; 25.98; 27.18;30.09; 30.34 (2×C); 30.65; 34.76; 42.32; 51.96; 53.96; 56.46; 58.36;61.24; 62.42; 62.69; 64.66; 69.02; 69.28; 70.20 (2×C); 70.66; 70.72(2×C); 70.92; 71.05; 73.05; 75.02; 75.86; 76.45; 76.78; 77.34; 77.68;99.89; 100.91; 102.55; 104.05; 112.25; 115.87; 122.22; 126.88; 148.86;151.60; 170.37; 174.79; 175.52 (2×C).

EXAMPLE B4.1 Preparation of Compound No. (22)

(a) 35 mg (52%) of disaccharide No. (24) are obtained according toExample B1.1(a) from 49 mg (100 μmol) of compound No. (23) and 78 mg(127 μmol) of UDP-gal (in this case the incubation mixture comprises 12%of DMSO (vol/vol)).

¹H-NMR (D₆-DMSO-CD₃OD-D₂O, 400.13 MHz) δ=1.18 (m, 8 H); 1.46 (m, 4 H);2.22 (t, 7.5 Hz, 2 H); 3.32-3.86 (m, 14 H); 3.58 (s, 3 H); 4.44 (d, 8.6Hz, 1 H); 6.12 (s, 1 H); remaining signals masked by the solvent.¹³C-NMR (D₆-DMSO-CD₃OD-D₂O, 62.89 MHz) δ=25.93; 26.98; 30.07; 30.24;30.29; 30.49; 35.00; 52.45; 57.55; 62.05; 62.73; 70.49; 71.80; 72.46;73.53; 74.99; 76.79; 76.91; 80.71; 100.81; 102.61; 105.20; 176.45; noresolution of the remaining signals.

(b) 41 mg (86%) of compound No. (25) are obtained according to Example B1.1(b) (in this case the buffer solution comprises 9% of DMSO (vol/vol))from 33 mg (51 μmol) of compound No. (24) and 53 mg (80 μmol) ofCMP-sia.

¹H-NMR (CD₃OD-D₂O, 250.13 MHz) δ=1.17 (m, 8 H); 1.45 (m, 4 H); 1.68(broad t, 11.0 Hz, 1 H); 1.94 (s, 3 H); 2.20 (t, 7.6 Hz, 2 H); 2.74(broad d, 11.0 Hz, 1 H); 3.29-4.02 (m, 24 H); 4.38 (d, 8.6 Hz, 1 H);4.42 (d, 8.6 Hz, 1 H); 6.05 (s, 1 H). ¹³C-NMR (CD₃OD-D₂O, 62.89 MHz)δ=22.35; 25.72; 26.93; 29.84; 30.06; 30.19; 30.29; 34.48; 41.62; 51.70;53.67; 57.00; 61.68; 62.44; 64.13; 68.87; 68.96; 69.02; 69.73; 70.58;72.70; 73.46; 74.65; 76.31; 76.74; 77.36; 80.93; 100.43; 100.90; 102.16;104.74; 166.82; 174.86; 175.26; 175.85; no resolution of the remainingsignals.

(c) 8 mg (62%) of compound No. (22) are obtained according to ExampleB1.1(c) from 11 mg (12 μmol) of compound No. (25) and 10 mg (60 μmol) ofGDP-D-arabinose. ¹H-NMR (CD₃OD, 500.00 MHz) δ=1.18 (m, 8 H); 1.48 (m, 4H); 1.66 (broad t, 11.0 Hz, 1 H); 1.95 (s, 3 H); 2.20 (t, 7.6 Hz, 2 H);2.72 (dd, 11.6 Hz, 2.8 Hz, 1 H); 3.36-4.03 (m, 28 H); 4.49 (d, 8.6 Hz, 2H); 4.58 (d, 8.6 Hz, 1 H); 5.02 (d, 3.6 Hz, 1 H); 6.09 (s, 1 H); ¹³C-NMR(CD₃OD, 126.00 MHz) δ=22.10; 25.55; 26.73; 29.67; 29.88; 29.96; 30.15;34.33; 41.53; 51.49; 53.48; 57.19; 60.82; 61.67; 62.50; 64.68; 68.42;68.85; 69.65; 69.67; 69.72; 70.34; 70.44; 70.47; 72.56; 74.53; 74.78;75.56; 76.28; 76.84; 77.40; 99.72; 100.42; 101.83; 103.29; 112.28;174.37; 175.01; 175.64; no resolution of the remaining signals.

EXAMPLE B4.2 Preparation of Compound No. (26)

7 mg (53%) of compound No. (26) are obtained according to ExampleB4.1(c) from 11 mg (12 μmol) of compound No. (25) and 11 mg (17 μmol) ofGDP-L-galactose. ¹H-NMR (CD₃OD, 500.00 MHz) δ=1.22 (m, 8 H); 1.48 (m, 4H); 1.67 (broad t, 11.0 Hz, 1 H); 1.97 (s, 3 H); 2.24 (t, 7.6 Hz, 2 H);2.84 (dd, 11.6 Hz, 2.8 Hz, 1 H); 3.38-4.08 (m, 29 H); 4.53 (broad d, 8.6Hz, 2 H); 4.69 (broad t, 5.5 Hz, 1 H); 5.03 (d, 3.6 Hz, 1 H); 6.13 (s, 1H); ¹³C-NMR (CD₃OD, 125.80 MHz) δ=22.58; 26.01; 27.23; 30.12; 30.33;30.40; 30.61; 34.79; 42.28; 51.96; 53.99; 57.85; 61.16; 62.20; 62.67;64.69; 69.08; 69.29; 70.13; 70.26; 70.81 (2×C); 70.91; 71.15; 71.39;73.02; 75.04; 75.79; 76.72; 76.99; 77.41; 77.72; 100.47; 100.96; 101.11;104.09; 112.21; 174.78; 175.51; 175.12; no resolution of the remainingsignals.

EXAMPLE B5.1 Preparation of Compound No. (48)

(a) 28 mg (83%) of compound No. (50) are obtained according to ExampleB1.1(a) (in this case the buffer solution comprises about 5% of DMSO(vol/vol)) from 24 mg (64 μmol) of No. (49) and 50 mg (80 μmol) ofUDP-gal.

¹H-NMR (CD₃OD-CDCl₃-D₂O, 250.13 MHz) δ=1.29 (m, 8 H); 1.58 (m, 4 H);2.30 (t, 7.6 Hz, 2 H); 8.13 (broad t, 8.5 Hz, 0.4 H); 3.36-3.98 (m, 16.6H); 4.40 (m, 2 H); 7.96 (s, 0.4 H); 8.15 (s, 0.6 H). M: main isomer; S:secondary isomer; ¹³C-NMR (CD₃OD-CDCl₃-D₂O, 62.90 MHz, DEPT) δ=25.54;26.42 M; 26.48 S; 29.60; 29.74; 29.79; 30.03; 34.53; 52.11; 55.10 M;59.69 S; 61.22; 62.03; 69.72; 70.67 M; 70.93 S; 72.11; 73.11 S; 73.40 M;74.05; 75.77 S; 75.93 M; 76.53; 79.96; 101.71 S; 101.90 M; 104.30;164.02 M; 167.98 S.

(b) 31 mg (77%) of compound No. (51) are obtained according to ExampleB1.1(b) from 26 mg (48 μmol) of compound No. (50) and 54 mg (64 μmol) ofCMP-sia.

¹H-NMR (CD₃OD, 250.13 MHz) δ=1.18 (m, 8 H); 1.56 (m, 4 H); 1.68 (t, 11.6Hz, 1 H); 2.00 (s, 3 H); 2.36 (t, 7.6 Hz, 2 H); 2.66 (dd, 11.6 Hz, 2.8Hz, 1 H); 3.49-4.02 (m, 23 H); 4.10 (dd, 11.08 Hz, 2.8 Hz, 1 H); 4.52(m, 2 H); 7.95 (s, 0.3 H); 8.16 (s, 0.7 H). ¹³C-NMR (CD₃OD, 62.90 MHz)δ=24.20; 26.44; 27.05; 30.21; 30.27; 30.41; 30.62; 35.87; 41.79; 53.84;54.21; 56.07 M; 60.92 S; 62.19; 63.19; 64.73; 69.62; 70.24; 70.52;71.55; 72.70 M; 73.09 S; 73.93; 74.39; 75.04; 76.93; 77.33; 77.63;80.37; 101.96; 102.61 S; 102.94 M; 104.73; 166.79 M; 170.26 S; 176.06;177.16; 180.09.

(c) 13 mg (29%) of compound No. (48) are obtained according to ExampleB1.1(c) from 37 mg (45 μmol) of compound No. (51) and 40 mg (61 μmol) ofGDP-L-galactose. ¹H-NMR (CD₃OD, 400.13 MHz) δ=1.25 (m, 8 H); 1.52 (m, 4H); 1.65 (broad t, 11.0 Hz, 1 H); 1.96 (s, 3 H); 2.25 (t, 7.6 Hz, 2 H);2.82 (dd, 11.6 Hz, 2.8 Hz, 1 H); 3.31-4.06 (m, 29 H); 4.26 (d, 8.6 Hz,0.4 H); 4.44 (d, 8.6 Hz, 0.6 H); 4.50 (m, 2 H); 5.08 (d, 4.3 Hz, 1 H);7.95 (s, 0.4 H); 8.15 (s, 0.6 H). ¹³C-NMR (CD₃OD, 100.60 MHz) δ=22.64;25.99; 26.99; 30.09; 30.24; 30.51; 34.78; 42.23; 51.98; 53.95; 56.11 M;60.86; 61.22; 62.01; 62.32; 62.64; 62.60; 64.57; 68.72; 68.98; 69.22;70.07; 70.14; 70.50; 70.53; 70.69; 70.87 (2×C); 70.96; 71.19; 73.05;74.97; 75.39 S; 75.65 M; 76.52 S; 76.65; 77.00; 77.28; 77.61; 100.12 M;100.27 S; 100.91; 102.11 M; 102.19; 103.95; 164.50 M; 168.49 S; 174.85;175.53; 176.05.

EXAMPLE B5.2 Preparation of Compound No. (64)

(a) 11 mg (29%) of disaccharide No. (65) are obtained as two amideisomers (about 60/40) according to Example B2.1(a) from 26 mg (69 μmol)of compound No. (49) and 52 mg (84 μmol) of UDP-gal.

¹H-NMR (CD₃OD-CDCl₃, 250.13 MHz) δ=1.22 (m, 8 H); 1.50 (m, 4 H); 2.22(t, 7.6 Hz, 2 H); 3.13-3.89 (m, 17 H); 4.22 (d, 8.6 Hz, 0.6 H); 4.24 (d,8.6 Hz, 0.4 H); 4.30 (d, 8.6 Hz, 0.4 H); 4.43 (d, 8.6 Hz, 0.6 H); 7.84(s, 0.4 H); 8.02 (s, 0.6 H. M: main isomer; S: secondary isomer; ¹³C-NMR(CD₃OD-CDCl₃, 62.90 MHz) δ=25.85; 26.81; 29.95; 30.10; 30.15; 30.41;34.76; 52.10; 55.20 M; 59.51 S; 62.34; 62.47; 70.00 M; 70.10 S; 70.32;70.70 M; 70.98 S; 77.07 S; 72.26 M; 74.33; 76.88; 76.98 S; 77.07 M;82.61 S, 83.99 M; 101.88 M; 102.07 S; 104.55 S; 105.01 M; 164.51 M;168.43 S; 176.21.

(b) 10 mg (55%) of compound No. (66) are obtained as two amide isomersaccording to Example B1.1(b) from 12 mg (22 μmol) of compound No. (65)and 23 mg (35 μmol) of CMP-sia.

¹H-NMR (CD₃OD, 250.13 MHz) δ=1.23 (m, 8 H); 1.48 (m, 4 H); 1.64 (broadt, 11.6 Hz, 1 H); 1.91 (s, 3 H); 2.21 (t, 7.6 Hz, 2 H); 2.77 (dd, 11.6Hz, 2.8 Hz, 1 H); 3.11-3.88 (m, 23 H); 3.93 (dd, 10.3 Hz, 3.4 Hz, 1 H);4.24 (d, 8.6 Hz, 0.4 H); 4.30 (d, 8.6 Hz, 0.6 H); 4.38 (d, 8.6 Hz, 0.4H); 4.44 (d, 8.6 Hz, 0.6 H); 7.83 (s, 0.4 H); 8.16 (s, 0.6 H). ¹³C-NMR(CD₃OD, 62.90 MHz) δ=22.61; 26.01; 26.99; 30.12; 30.27; 30.33; 30.59;34.79; 42.22; 51.98; 53.90; 55.32 M; 59.94 S; 62.70 (2×C); 65.00; 69.09;69.34; 70.29; 7.50; 70.64 M; 70.96 S; 73.00; 74.91; 77.06; 77.42; 77.57(2×C); 83.00 S; 84.08 S; 101.17; 102.15 M; 102.63 S; 105.11 S; 105.19 M;164.71 M; 168.49 S; 174.93; 175.49; 176.05.

(b′) Alternatively, the enzyme reactions (a) and (b) can also be carriedout together in one reaction step. 9 mg (37%) of trisacchande No. (66)are thus obtained from 10 mg (27 μmol) of amide No. (49), 31 mg (51μmol) of UDP-gal and 33 mg (51 μmol) of CMP-sia.

(c) 10 mg (89%) of compound No. (64) are obtained according to ExampleB2.1(c) from 10 mg (12 μmol) of compound No. (66) and 10 mg (15 μmol) ofGDP-D-arabinose. ¹H-NMR (CD₃OD, 400.13 MHz) δ=1.22 (m, 8 H); 1.50 (m, 4H); 1.61 (broad t, 11.0 Hz, 1 H); 1.92 (s, 3 H); 2.23 (t, 7.6 Hz, 2 H);2.79 (dd, 11.6 Hz, 2.8 Hz, 1 H); 3.23-4.04 (m, 29 H); 4.22 (d, 8.6 Hz,0.4 H); 4.46 (m, 1.2 H); 4.63 (d, 8.6 Hz, 0.4 H); 5.00 (d, 4.3 Hz, 1 H);7.84 (s, 0.4 H); 8.09 (s, 0.6 H). ¹³C-NMR (CD₃OD, 62.90 MHz) δ=22.59;26.01; 27.02; 30.12; 30.28 (2×C); 30.61; 34.79; 42.48; 51.99; 53.88;56.30 H; 61.30; 62.95; 63.06; 64.77; 65.13; 65.32; 68.63; 69.25; 70.28(2×C); 70.43; 70.67; 71.05; 73.05; 73.52; 74.05; 74.94; 76.65; 76.91;77.27; 77.53; 77.0; 100.10 H; 100.19 N; 100.75; 100.89; 102.03; 102.29;104.01; 104.34; 164.90 H; 168.31 N; 174.60; 174.83; 175.39; 175.51;176.55.

EXAMPLE B5.3 Preparation of Compound No. (67)

11 mg (about 73%) of compound No. (67) are obtained according to ExampleB5.2(c) from 13 mg (15 μmol) of compound No. (66) and 11 mg (18 μmol) ofGDP-2-fluoro-fucose. The material isolated comprises about 80% ofproduct No. (67). ¹H-NMR (CD₃OD, 400.13 MHz) δ=1.09 (d, 6.8 Hz, 3 H);1.61 (broad t, 11.0 Hz, 1 H); 1.91 (s, 3 H); 2.22 (t, 7.6 Hz, 2 H); 2.79(dd, 11.6 Hz, 2.8 Hz, 1 H); 4.20-4.59 (several d, 3 H); 4.82 (broad q,6.8 Hz, 0.8 H); 5.15 (d, 4.3 Hz, 0.8 H). ¹³C-NMR (CD₃OD, 62.90 MHz)δ=16.40; 34.79; 91.01 (d, 180 Hz). ¹⁹F-NMR (CD₃OD, 376.5 MHz) δ=−211.4;−210.95; ratio of the two signals: about 70/30.

EXAMPLE B6.1 Preparation of Compound No. (77)

(a) 9 mg (34%) of compound No. (79) are obtained according to ExampleB2.1(a) from 20 mg (49 μmol) of compound No. (78) and 36 mg (58 μmol) ofUDP-gal (in this case the incubation mixture comprises 5% of DMSO(vol/vol)).

¹H-NMR (CD₃OD, 250.13 MHz) δ=1.16 (t, 7.5 Hz, 3 H); 1.22 (m, 8 H); 1.49(m, 4 H); 2.22 (t, 7.6 Hz, 2 H); 3.15-3.88 (m, 17 H); 3.99 (q, 7.5 Hz, 2H); 4.25 (d, 8.6 Hz, 1 H); 4.31 (d, 8.6 Hz, 1 H). ¹³C-NMR (CD₃OD, 100.61MHz) δ=15.04; 26.03; 27.02; 30.14; 30.31; 30.38; 30.60; 34.78; 51.97;57.85; 61.99; 62.52; 62.70; 70.22; 70.55; 70.73; 72.44; 74.53; 77.11;77.44; 84.71; 102.78; 105.27; 159.49; 176.01.

(b) 18 mg (78%) of compound No. (80) are obtained according to ExampleB1.1(b) from 16 mg (27 μmol) of compound No. (79) and 25 mg (37 μmol) ofCMP-sia (in this case the incubation mixture comprises 8% of DMSO(vol/vol)).

¹H-NMR (CD₃OD, 250.13 MHz) δ=1.17 (t, 7.5 Hz, 3 H); 1.20 (m, 8 H); 1.48(m, 4 H); 1.68 ((broad t, 11.6 Hz, 1 H); 1.91 (s, 3 H); 2.21 (t, 7.6 Hz,2 H); 2.73 (dd, 11.6 Hz, 2.8 Hz, 1 H; 3.16-4.08 (m, 25 H); 4.30 (d, 8.6Hz, 1 H); 4.33 (broad d, 8.6 Hz, 1 H). ¹³C-NMR (CD₃OD, 62.98 MHz)δ=15.19; 22.66; 26.03; 27.00; 30.13; 30.31; 30.37; 30.61; 34.78; 42.00;51.98; 53.94; 57.69; 62.00; 62.74 (2×C); 69.10; 69.36; 69.56; 69.87;70.48 (2×C); 70.73; 72.87; 74.88; 76.84; 77.42 (2×C); 84.41; 101.30;102.99; 104.55; 159.32; 175.45 (2×C); 175.54.

(b′) Steps (b) and (c) can also be carried out as a one-pot reactionaccording to Example B5.2(b). 9 mg (38%) of compound No. (80) areobtained from 12 mg (28 μmol) of compound No. (78), 20 mg (33 μmol) ofUDP-gal and 24 mg (36 μmol) of CMP-sia.

(c) 10 mg (94%) of compound No. (77) are obtained according to ExampleB2.1(c) from 9 mg (11 μmol) of compound No. (80) and 12 mg (19 μmol) ofGDP-L-galactose. ¹H-NMR (CD₃OD, 400.13 MHz) δ=1.17 (t, 7.5 Hz, 3 H);1.21 (m, 8 H); 1.45 (m, 4 H); 1.65 (broad t, 12.4 Hz, 1 H); 1.91 (s, 3H); 2.21 (t, 8.4 Hz, 2 H); 2.73 (dd, 12.4 Hz, 3.4 Hz, 1 H); 3.21-4.16(m, 31 H); 4.39-4.59 (m, 2 H); 4.65 (broad t, 7.0 Hz, 1 H); 4.99 (d, 4.8Hz, 1 H). ¹³C-NMR (CD₃OD, 126.00 MHz) δ=15.53; 22.61; 26.03; 27.02;30.13; 30.31; 30.36; 30.65; 34.79; 41.96; 51.98; 53.93; 59.40; 61.36;62.05; 62.74; 64.25; 68.87; 69.40; 69.79; 70.33; 70.72 (2×C); 70.89(2×C); 71.17; 72.95; 74.38; 74.90; 76.48; 77.17 (2×C); 77.22; 77.39;99.85; 101.30; 102.34; 104.35; 158.96; 175.08; 175.45; 176.02.

EXAMPLE B6.2 Preparation of Compound No. (81)

5 mg (48%) of compound No. (81) are obtained according to ExampleB6.1(c) from 9 mg (11 μmol) of compound No. (80) and 11 mg (17 μmol) ofGDP-L-glucose. ¹H-NMR (CD₃OD, 250.13 MHz) δ=1.28 (t, 7.5 Hz, 3 H); 1.32(m, 8 H); 1.49 (m, 4 H); 1.76 (broad t, 12.4 Hz, 1 H); 2.02 (s, 3 H);2.31 (t, 8.4 Hz, 2 H); 2.80 (dd, 12.4 Hz, 3.4 Hz, 1 H); 3.19-4.28 (m, 31H); 4.51 (m, 3 H); 5.09 (d, 4.8 Hz, 1 H). ¹³C-NMR (CD₃OD, 126.00 MHz)δ=15.53; 22.59; 26.03; 27.02; 30.14; 30.31; 30.36; 30.64; 34.79; 41.62;51.96; 53.89; 58.97; 61.45; 61.79; 62.88; 64.10; 69.23; 69.57; 70.75;71.02 (2×C); 71.88; 72.70; 73.16; 73.76; 74.61; 74.63; 76.76 (2×C);77.22 (2×C); 77.43; 101.02; 102.48; 104.36; 105.13; 175.41; noresolution of the remaining signals.

EXAMPLE B7 Preparation of Compound No. (93)

(a) 13 mg (61%) of disaccharide No. (96) are obtained according toExample B2.1(a) from 15 mg (37 μmol) (in this case the buffer solutioncomprises 8% of DMSO (vol/vol)) of compound No. (95) and 34 mg (56 μmol)of UDP-D-galactose.

¹H-NMR (CD₃OD-D₂O-CDCl₃, 400.13 MHz) δ=1.22 (m, 8 H); 1.49 (m, 4 H);2.26 (t, 7.5 Hz, 2 H); 3.27-3.84 (m, 17 H); 3.96 (q, 15.2 Hz, 2 H); 4.28(d, 8.6 Hz, 1 H); 4.52 (d, 8.6 Hz, 1 H). ¹³C-NMR (CD₃OD-D₂O-CDCl₃, 100.6MHz) DEPT δ=25.76; 26.62; 29.78; 29.93; 29.96; 30.22; 34.76; 52.49;55.75; 62.28; 62.33; 62.45; 69.97; 70.28; 71.04; 72.15; 74.08; 76.72;77.01; 83.54; 101.94; 104.87.

(b) 14 mg (79%) of compound No. (97) are obtained according to ExampleB1.1(b) from 12 mg (21 μmol) of compound No. (96) and 18 mg (29 μmol) ofCMP-sia.

¹H-NMR (CD₃OD, 400.13 MHz) δ=1.19-1.32 (m, 8 H); 1.44-1.57 (m, 4 H);1.63 (t, 11.6 Hz, 1 H); 1.95 (s, 3 H); 2.25 (t, 7.6 Hz, 2 H); 2.80 (dd,11.6 Hz, 2.8 Hz, 1 H); 3.27 (m, 1 H); 3.33-3.85 (m, 22 H); 3.96 (dd, 3.7Hz, 9.8 Hz, 1 H); 3.98 (q, 16.6 Hz, 2 H); 4.28 (d, 8.6 Hz, 1 H); 4.50(d, 8.6 Hz, 1 H). ¹³C-NMR (CD₃OD, 100.6 MHz) δ=22.61; 26.01; 26.98;30.11; 30.28; 30.32; 30.60; 34.79; 42.32; 51.64; 53.91; 55.63; 62.09;62.68; 62.73; 62.82; 64.78; 68.80; 69.27; 70.16; 70.53; 70.62; 71.35;73.22; 74.84; 77.03; 77.56; 84.49; 100.94; 102.22; 105.35; 174.75;175.49; 176.02; 176.07.

(c) 14 mg (100%) of compound No. (93) are obtained according to ExampleB2.1(c) from 12 mg (14 μmol) of compound No. (97) and 14 mg (22 μmol) ofGDP-D-arabinose. ¹H-NMR (CD₃OD, 400.13 MHz) δ=1.14-1.28 (m, 8 H);1.35-1.50 (m, 4 H); 1.56 (t, 12.4 Hz, 1 H); 1.89 (s, 3 H); 2.19 (t, 8.4Hz, 2 H); 2.74 (dd, 12.4 Hz, 3.4 Hz, 1 H); 3.23-3.89 (m, 27 H); 3.99 (m,2 H); 4.33 (d, 7.0 Hz, 1 H); 4.42 (d, 8.6 Hz, 1 H); 4.61 (broad d, 12.9Hz, 2 H); 4.96 (d, 4.3 Hz, 1 H). ¹³C-NMR (CD₃OD, 100.6 MHz) δ=22.61;26.02; 26.99; 30.11; 30.30; 30.33; 30.62; 34.79; 42.55; 51.99; 53.84;56.81; 61.38; 62.13; 62.84; 63.03; 64.40; 64.82; 65.36; 68.29; 69.37;70.29; 70.66; 71.03; 71.40; 73.17; 73.63; 74.22; 74.78; 77.35; 77.64;77.80; 100.33; 100.88; 102.15; 104.29; 174.71; 175.44; 175.94; 176.09.

EXAMPLE B8.1 Preparation of Compound No. (52)

(a) 8.7 g (17.0 mmol) of benzyl-protected monosaccharide No. (54) areinitially introduced into the reaction vessel together with 5.5 g (22mmol) of mercury cyanide in 260 ml of dry toluene/nitromethane(vol/vol—1/1) and the mixture is stirred with triturated, activemolecular sieve 4 Å (about 5 g) at RT for 30 minutes. 10.3 g (25.0 mmol)of per-O-acetylated α-galactosyl bromide, dissolved in 35 ml oftoluene/nitromethane (see above) are then added drop-wise to thismixture and the mixture is heated at 50° C. for about 18 hours. Afterall the monosaccharide has reacted, the mixture is filtered carefullyover Celite, the solvent is removed in a rotary evaporator and theresidue which remains is chromatographed over silica gel (eluent:hexane/ethyl acetate—2/1). 9.1 g (64%) of disaccharide No. (55) areobtained.

¹H-NMR (CDCl₃, 400.13 MHz) δ=1.22 (m, 8 H); 1.51 (m, 4 H); 1.88 (s, 3H); 1.89 (s, 3 H); 1.91 (s, 3 H); 2.05 (s, 3 H); 2.23 (t, 7.6 Hz, 2 H);3.06 (broad, 1 H); 3.41 (m, 2 H); 3.59 (m, 5 H); 3.71 (m, 2 H); 3.78(dt, 6.1 Hz, 9.1 Hz, 1 H); 3.98 (dd, 6.6 Hz, 11.4 Hz, 1 H); 4.25 (dd,6.1 Hz, 11.4 Hz, 1 H); 4.39 (m, 1 H); 4.50 (m, 2 H); 4.59 (d, 7.3 Hz, 1H); 4.89 (dd, 3.6 Hz, 10.9 Hz, 1 H); 5.05 (m, 1 H); 5.13 (dd, 7.3 Hz,10.9 Hz, 1 H); 5.19 (dq, 1.2 Hz, 11.5 Hz, 1 H); 5.22 (dd, 0.6 Hz, 3.0Hz, 1 H); 5.27 (m, 1 H); 5.47 (s, 1 H); 5.86 (m, 1 H); 7.30 (m, 3 H);7.40 (m, 2 H). ¹³C-NMR (CDCl₃, 62.90 MHz) δ=20.52 (2×C); 20.62 (2×C);24.80; 25.65; 28.93; 29.0 (2×C); 29.38; 33.99; 51.44; 58.08; 60.70;65.60; 65.87; 66.73; 68.70; 69.06; 70.27; 70.40; 70.97; 76.49; 78.63;80.18; 101.01; 101.33; 117.88; 126.03 (2×C); 128.15 (2×C); 129.14;132.44; 137.04; 155.43; 169.40; 170.06; 170.11; 170.24; 174.42.

(b) 9.1 g (10.7 mmol) of disaccharide No. (55) are dissolved in 100 mlof methylene chloride, and 5 ml of a 90% trifluoroacetic acid are addedat room temperature. After about 6 hours, the mixture is neutralizedwith saturated sodium bicarbonate solution, diluted with ethyl acetateand extracted in succession with water and saturated sodium chloridesolution. The organic phase is dried over sodium sulfate and evaporated.7 ml of pyridine and 3.5 ml of acetic anhydride are added to theresulting residue and the mixture is stirred overnight at RT. Themixture is then diluted with ethyl acetate and extracted successivelywith 4 N hydrochloric acid, water and saturated sodium bicarbonatesolution. After evaporation of the solvent, a yellow syrup remains, andis chromatographed over silica gel (eluent: petroleum ether/ethylacetate—2/1). 6.9 g (76%) of disaccharide No. (56) are obtained.

¹H-NMR (CDCl₃, 400.13 MHz) δ=1.22 (m, 8 H); 1.51 (m, 4 H); 1.93 (s, 3H); 1.98 (s, 3 H); 2.00 (s, 3 H); 2.01 (s, 3 H); 2.09 (s, 3 H); 2.17 (s,3 H); 2.24 (t, 7.6 Hz, 2 H); 3.10 (m, 1 H); 3.39 (dt, 6.0 Hz, 10.9 Hz, 1H); 3.58 (m, 1 H); 3.60 (s, 3 H); 3.79 (m, 2 H); 4.04 (m, 3 H); 4.17(dd, 6.0 Hz, 11.0 Hz, 1 H); 4.80 (m, 1 H); 4.52 (m, 3 H); 4.66 (m, 1 H);4.88 (m, 2 H); 4.99 (m, 1 H); 5.01 (dd, 7.3 Hz, 11.5 Hz, 1 H); 5.19 (dq,0.6 Hz, 12.1 Hz, 1 H); 5.28 (m, 2 H); 5.27 (m, 1 H); 5.90 (m, 1 H).¹³C-NMR (CDCl₃, 62.90 MHz) δ=20.50; 20.61 (3×C); 20.67; 20.79; 24.79;25.63; 28.91; 28.97; 29.01; 29.30; 33.99; 51.43; 58.02; 60.98; 62.44;65.59; 66.76 (2×C); 69.00; 69.15; 70.00; 70.42; 70.95; 71.65; 100.55;101.02; 117.91; 137.50; 155.55; 169.15; 169.27; 170.11; 170.19; 170.32;170.75; 174.29.

(c) 4.0 g (4.7 mmol) of disaccharide No. (56) are dissolved in 60 ml ofabsolute THF under argon at RT and 5.6 ml of diethyl malonate and 0.4 g(0.3 mmol) of tetrakis-(triphenyl)-palladium are added in succession.After about 1 hour, the solvent is evaporated off and the residue whichremains is chromatographed over silica gel. 3.1 g (89%) of amine No.(57) are obtained.

¹H-NMR (CDCl₃, 250.13 MHz) δ=1.33 (m, 8 H); 1.60 (m, 4 H); 1.99 (s, 3H); 2.05 (m, 12); 2.13 (s, 3 H); 2.29 (t, 7.6 Hz, 2 H); 2.92 (dd, 7.5Hz, 8.2 Hz, 1 H); 3.46 (dt, 6.9 Hz, 10.3 Hz, 1 H); 3.58 (m, 1 H); 3.67(s, 3 H); 3.89 (m, 2 H); 4.14 (m, 6 H); 4.73 (d, 7.6 Hz, 1 H); 4.99 (m,2 H); 5.15 (dd, 7.6 Hz, 11.7 Hz, 1 H); 5.35 (m, 1 H). ¹³C-NMR (CDCl₃,62.90 MHz) δ=20.50; 20.60 (3×C); 20.77; 20.81; 24.82; 25.83; 28.98;29.09 (2×C); 29.42; 33.99; 51.40; 57.05; 60.91; 62.51; 66.74; 68.70;69.52; 70.16; 70.58; 70.97; 72.04; 83.53; 101.45; 103.12; 169.03;169.30; 170.13; 170.29; 170.75; 174.44.

(d) 287 mg (98%) of amide are obtained according to Example A8 from 56mg (360 μmol) of 3,5-dihydroxybenzoic acid and 250 mg (330 μmol) ofamine No. (57) in the presence of 155 mg of HBPyU, after chromatographyof the reaction mixture over silica gel (eluent: methylenechloride/methanol—15/0.5), and the product is immediately deacetylatedwith sodium methanolate. Renewed chromatography over silica gel (eluent:methylene chloride/methanol/water—6/4/1) gives 137 mg (65%) ofdisaccharide No. (58).

¹H-NMR (CD₃OD 400.13 MHz) δ=1.08 (m, 8 H); 1.41 (m, 4 H); 2.18 (t, 7.6Hz, 2 H); 3.19-3.89 (m, 17 H); 4.22 (d, 8.6 Hz, 1 H); 4.56 (broad d, 9.0Hz, 1 H); 6.30 (t, about 2.0 Hz, 1 H); 6.64 (d, about 2.0 Hz, 2 H).¹³C-NMR (CD₃OD, 100.61 MHz) δ=26.53; 27.13; 30.04; 30.23; 30.30; 30.61;34.76; 51.95; 56.90; 62.39; 62.71; 70.14; 70.69; 70.77; 72.32; 74.36;76.98; 77.45; 84.24; 102.33; 105.21; 106.57; 107.03 (2×C); 138.03;159.70 (2×C); 171.39; 176.19.

(e) 43 mg (87%) of compound No. (59) are obtained according to ExampleB1.1(b) from 34 mg (53 μmol) of compound No. (58) and 50 mg (75 μmol) ofCMP-sia.

¹H-NMR (CD₃OD, 400.13 MHz) δ=1.12 (m, 8 H); 1.46 (m, 4 H); 1.72 (broadt, 11.6 Hz, 1 H); 1.98 (s, 3 H); 2.22 (t, 7.6 Hz, 2 H); 2.74 (dd, 11.6Hz, 2.8 Hz, 1 H); 3.33 (m, 1 H); 3.42-3.75 (m, 16 H); 3.83-3.97 (m, 7H); 4.38 (d, about 8.6 Hz, 1 H); 4.59 (broad d, about 8.6 Hz, 1 H); 6.37(t, about 2.0 Hz, 1 H); 6.68 (d, about 2.0 Hz, 2 H). ¹³C-NMR (CD₃OD,100.61 MHz) δ=22.74; 25.99; 27.14; 30.09; 30.30; 30.36; 30.67; 34.80;41.40; 51.97; 53.91; 56.71; 62.61; 62.78; 63.96; 69.34; 69.71; 70.67;70.78; 70.83; 72.89; 74.85; 76.68; 77.30; 77.45; 82.95; 101.56; 102.56;104.07; 106.74; 107.70 (2×C); 138.23; 159.71; 171.33; 175.21; 175.48;176.23.

(f) 10 mg (72%) of compound No. (52) are obtained according to ExampleB2.1(c) from 12 mg (13 μmol) of compound No. (59) and 14 mg (17 μmol) ofGDP-D-arabinose. ¹H-NMR (CD₃OD, 400.13 MHz) δ=1.10 (m, 8 H); 1.41 (m, 4H); 1.65 (broad t, 11.0 Hz, 1 H); 1.92 (s, 3 H); 2.18 (t, 7.6 Hz, 2 H);2.72 (dd, 2.8 Hz, 11.6 Hz, 1 H); 3.33-3.90 (m, 28 H); 4.37 (broad t, 6.3Hz, 1 H); 4.48 (d, 8.6 Hz, 1 H); 4.56 (d, 8.6 Hz, 1 H); 5.01 (d, 4.3 Hz,1 H); 6.35 (t, about 3.0 Hz, 1 H); 6.64 (d, about 3 Hz, 2 H). ¹³C-NMR(CD₃OD, 100.61 MHz) δ=22.68; 25.96; 27.14; 30.03; 30.19; 30.30; 30.62;34.83; 41.91; 52.15; 53.81; 58.37; 61.45; 62.98; 64.10; 65.50; 68.88;69.42; 69.66; 70.24; 70.33; 70.88; 70.94; 71.00; 72.84; 74.85 (2×C);76.15; 76.43; 77.11; 77.76; 100.00; 100.34; 101.52; 101.95; 103.27;106.92 (2×C); 138.15; 159.70 (2×C); 171.31; 174.84; 175.66; 176.62.

EXAMPLE B8.2 Preparation of Compound No. (60)

8 mg (32%) of compound No. (60) are obtained according to ExampleB8.1(f) from 23 mg (24 μmol) of compound No. (59) and 17 mg (28 μmol) ofGDP-2-amino-fucose. ¹H-NMR (CD₃OD, 400.13 MHz) δ=1.16 (m, 11 H); 1.46(m, 4 H); 1.68 (broad t, 11.0 Hz, 1 H); 1.96 (s, 3 H); 2.22 (t, 7.6 Hz,2 H); 2.77 (dd, 2.8 Hz, 11.6 Hz, 1 H); 3.23 (dd, 5.5 Hz, 12.4 Hz, 1 H);3.34 (m, 1 H); 3.39-3.95 (m, 24 H); 4.50-4.66 (m, 3 H); 5.26 (d, 4.3 Hz,1 H); 6.35 (t, about 3.0 Hz, 1 H); 6.66 (d, about 3 Hz, 2 H). ¹³C-NMR(CD₃OD, 100.61 MHz) δ=16.66; 22.61; 26.01; 27.21; 30.10; 30.28; 30.71;34.81; 41.90; 51.97; 52.50; 53.88; 59.21; 62.66; 63.04; 64.51; 68.14;68.55; 69.15; 69.44; 69.91; 70.81; 71.03; 72.55; 73.00; 73.97; 77.02(2×C); 76.58 (2×C); 77.98; 95.46; 101.40; 101.82; 103.19; 106.84 (2×C);106.97; 138.25; 159.89 (2×C); 171.46; 174.78; 175.53; 176.24.

EXAMPLE B8.3 Preparation of Compound No. (68)

(a) 66 mg of 3,5-di-O-acetoxybenzyl alcohol are cooled to 0° C. in 3 mlof dry toluene under argon. 1.6 ml of a 20% phosgene solution (toluene)are then added to this mixture and the mixture is stirred at RT forabout 2.5 hours and then evaporated to dryness. 250 mg (327 μmol) ofamine No. (57), dissolved in 3 ml of dry DMF, are immediately added tothis residue, 50 ml (1.2 equivalents) of triethylamine are added and themixture is stirred overnight at RT. Thereafter, the solvent isevaporated off and the residue is chromatographed over silica gel(eluent: methylene chloride/methanol—10/0.4). 229 mg (77%) ofperacetylated adduct are obtained, and are immediately dissolved in 5 mlof dry methanol, and 0.5 ml of a freshly prepared 0.1% sodiummethanolate solution is added. Customary working up gives 37 mg (25%) ofcarbamate No. (69).

¹H-NMR (CD₃OD, 400.13 MHz) δ=1.28 (m, 8 H); 1.51 (m, 4 H); 2.24 (t, 7.5Hz, 2 H); 3.24-3.86 (m, 17 H); 4.33 (d, 8.6 Hz, 1 H); 4.44 (broad d, 8.6Hz, 1 H); 4.91 (m, 2 H); 6.15 (t, about 2.0 Hz, 1 H); 6.27 (d, about 2.0Hz, 2 H). ¹³C-NMR (CD₃OD, 100.60 MHz) δ=25.99; 26.95; 30.10; 30.23;30.30; 30.53; 34.80; 51.98; 58.13; 62.53; 62.70; 67.46; 70.29; 70.59;70.86; 72.56; 74.44; 76.99; 77.42; 84.14; 102.64; 102.98; 105.09; 106.96(2×C); 140.40; 159.61 (2×C); 176.17; no resolution of the remainingsignals.

(b) 32 mg (61%) of compound No. (70) are obtained according to ExampleB1.1(b) from 37 mg (55 μmol) of compound No. (69) and 48 mg (73 μmol) ofCMP-sia.

¹H-NMR (CD₃OD, 400.13 MHz) δ=1.24 (m, 8 H); 1.51 (m, 4 H); 1.76 (broadt, 11.6 Hz, 1 H); 1.98 (s, 3 H); 2.25 (t, 7.6 Hz, 2 H); 2.80 (dd, 11.6Hz, 2.8 Hz, 1 H); 3.34-4.07 (m, 24 H); 4.43 (m, 2 H); 4.98 (m, 2 H);6.16 (t, about 2.0 Hz, 1 H); 6.29 (d, about 2.0 Hz, 2 H); ¹³C-NMR(CD₃OD, 100.61 MHz) δ=22.71; 25.99; 26.95; 30.11; 30.24; 30.29; 30.53;34.80; 41.79; 51.99; 53.93; 58.22; 62.74 (2×C); 64.29; 67.82; 68.11;69.27; 69.35; 69.92; 70.50; 70.86 (2×C); 72.61; 74.87; 76.67; 77.38(2×C); 83.79; 101.19; 102.54; 103.03; 104.49; 107.06 (2×C); 140.42;159.55 (2×C); 175.51; 176.20; no resolution of the remaining signals.

(c) 9 mg (64%) of compound No. (68) are obtained according to ExampleB2.1(c) from 12 mg (12 μmol) of compound No. (70) and 11 mg (17 μmol) ofGDP-D-arabinose. ¹H-NMR (CD₃OD, 400.13 MHz) δ=1.19 (m, 8 H); 1.46 (m, 4H); 1.65 (broad t, 11.0 Hz, 1 H); 1.92 (s, 3 H); 2.20 (t, 7.6 Hz, 2 H);2.78 (dd, 2.8 Hz, 11.6 Hz, 1 H); 3.22-3.94 (m, 28 H); 4.09 (broad t, 8.8Hz, 1 H); 4.39 (broad d, 8.6 Hz, 1 H); 4.56 (broad d, 8.6 Hz, 1 H); 4.98(m, 3 H); 6.09 (t, about 3.0 Hz, 1 H); 6.28 (d, about 3 Hz, 2 H).¹³C-NMR (CD₃OD, 100.61 MHz) δ=22.61; 26.01; 26.96; 30.12; 30.25; 30.29;30.56; 34.82; 42.09; 51.98; 53.95; 61.54; 63.25; 64.30; 65.70; 67.62;68.12; 69.18; 69.48; 69.90; 70.38; 70.84; 71.03 (2×C); 72.66; 74.52;74.97; 76.17; 77.00; 77.15; 77.66; 100.21; 101.31; 102.18; 103.34;103.68; 107.72 (2×C); 140.58; 159.60 (2×C); 175.15; 175.47; 176.21; noresolution of the remaining signals.

EXAMPLE B8.4 Preparation of Compound No. (71)

(a) 550 mg of peracetylated compound No. (56) are dissolved in 25 ml ofdry methanol and the solution is treated with 0.5 ml of a 0.1% sodiummethanolate solution. After about 1 h at RT, the mixture is neutralizedwith DOWEX 50×8 (H⁺ form) and filtered and the solvent is evaporated.Chromatography of the residue over silica gel (eluent: methylenechloride/methanol—9/1) gives 350 mg (82%) of disaccharide No. (72).

¹H-NMR (CD₃OD, 250.13 MHz) δ=1.18 (m, 8 H); 1.45 (m, 4 H); 2.19 (t, 7.6Hz, 2 H); 3.14-3.84 (m, 17 H); 4.32 (d, 8.6 Hz, 1 H); 4.42 (m, 2 H);5.05 (m, 1 H); 5.80 (m, 1 H). ¹³C-NMR (CD₃OD, 62.90 MHz) δ=25.97; 26.94;30.05; 30.22; 30.27; 30.51; 34.80; 52.17; 58.48; 62.58 (2×C); 66.70;70.14 H; 70.49; 70.88; 72.66; 74.41; 76.99; 77.27; 84.64; 102.98;105.10; 117.62; 134.34; 159.73; 176.41.

(b) 36 mg (71%) of compound No. (73) are obtained according to ExampleB1.1(b) from 34 mg (56 μmol) of compound No. (72) and 49 mg (74 μmol) ofCMP-sia.

¹H-NMR (CD₃OD, 250.13 MHz) δ=1.23 (m, 8 H); 1.49 (m, 4 H); 1.67 (broadt, 11.6 Hz, 1 H); 1.93 (s, 3 H); 2.22 (t, 7.6 Hz, 2 H); 2.75 (dd, 11.6Hz, 2.8 Hz, 1 H); 3.16-3.90 (m, 23 H); 3.94 (dd, 10.3 Hz, 3.4 Hz, 1 H);4.30-4.62 (m, 4 H); 5.10 (m, 1 H); 5.23 (m, 1 H); 5.86 (m, 1 H). ¹³C-NMR(CD₃OD, 62.98 MHz) δ=22.68; 26.02; 27.01; 30.12; 30.30; 30.35; 30.60;34.78; 41.77; 51.98; 53.95; 57.74; 62.71 (2×C); 64.25; 66.59; 69.09;69.32; 69.83; 70.46; 70.76; 72.84; 74.89; 76.86; 77.40 (3×C); 83.91;101.23; 102.91; 104.54; 117.43; 134.65; 158.92; 175.47; 176.02; 176.53.

(c) 18 mg (82%) of compound No. (71) are obtained according to ExampleB2.1(c) from 19 mg (22 μmol) of compound No. (73) and 18 mg (29 μmol) ofGDP-D-arabinose. ¹H-NMR (CD₃OD, 250.13 MHz) δ=1.21 (m, 8 H); 1.49 (m, 4H); 1.67 (broad t, 12.4 Hz, 1 H); 1.93 (s, 3 H); 2.24 (t, 8.4 Hz, 2 H);2.76 (dd, 12.4 Hz, 3.4 Hz, 1 H); 3.24-3.96 (m, 22 H); 4.08 (broad t, 6.4Hz, 1 H); 4.33-4.70 (m, 5 H); 5.00 (d, 4.8 Hz, 1 H); 5.13 (m, 1 H); 5.25(m, 1 H); 5.89 (m, 1 H). ¹³C-NMR (CD₃OD, 62.90 MHz) δ=22.68; 25.98;26.95; 30.06; 30.26; 30.57; 30.89; 34.82; 42.22; 52.15; 53.87; 59.71;61.45; 63.03; 64.49; 65.32; 66.91; 68.08; 68.64; 69.78; 70.17; 70.27;70.92 (2×C); 73.15; 74.79; 74.80; 76.42; 77.12; 77.51; 100.60; 101.56;102.85; 104.15; 118.21; 134.61; 175.04; 175.63; 176.21; no resolution ofthe remaining signals.

EXAMPLE B8.5 Preparation of Compound No. (82)

27 mg (77%) of compound No. (82) are obtained according to ExampleB8.4(c) from 30 mG (34 μmol) of compound No. (73) and 31 mg (47 μmol) ofGDP-L-galactose. ¹H-NMR (CD₃OD, 400.13 MHz) δ=1.15-1.33 (m, 8 H);1.40-1.56 (m, 4 H); 1.68 (t, 12.4 Hz, 1 H); 1.94 (s, 3 H); 2.23 (t, 8.4Hz, 2 H); 2.72 (dd, 12.4 Hz, 3.4 Hz, 1 H); 3.28-3.96 (m, 29 H); 4.03(broad t, 8.8 Hz, 1 H); 4.35-4.71 (m, 4 H); 4.99 (d, 4.8 Hz, 1 H); 5.12(m, 1 H); 5.25 (m, 1 H); 5.88 (m, 1 H). ¹³C-NMR (CD₃OD, 100.6 MHz)δ=22.67; 26.01; 27.02; 30.12; 30.32 (2×C); 30.64; 34.79; 41.95; 51.99;53.97; 59.37; 61.33; 62.45; 62.70; 64.13; 66.60; 68.80; 69.29; 69.71;70.28; 70.78 (2×C); 70.86; 71.10 (2×C); 72.65; 74.22; 74.90; 76.44;77.17; 77.37; 78.37; 99.81; 101.31; 102.37; 104.28; 117.71; 134.77;158.80; 175.10; 175.53; 176.04.

EXAMPLE B8.6 Preparation of Compound No. (98)

(a) 4.08 g (6.8 mmol) of disaccharide No. (72) are dissolved in asolvent mixture of THF (160 ml) and methanol (60 ml) at RT, and 0.50 g(1.2 mmol) of DPPB, 1.5 g (11.3 mmol) of sodium thiophenolate and 0.30 g(0.3 mmol) of Pd(dba)₂ (Aldrich) are added in succession under an argonatmosphere. After the mixture has been stirred overnight, the solventsare evaporated off and the residue is chromatographed over silica gel(eluent: methylene chloride/methanol—5/1). 2.23 g (64%) of amine No.(99) are obtained.

¹H-NMR (CD₃OD-CDCl₃, 400.13 MHz) δ=1.29-1.44 (m, 8 H); 1.56-1.69 (m, 4H); 2.33 (t, 7.5 Hz, 2 H); 2.80 (broad t, 7.4 Hz, 1 H); 3.35 (m, 1 H);3.44-3.96 (m, 15 H); 4.31 (d, 8.6 Hz, 1 H); 4.43 (d, 8.6 Hz, 1 H).

(b) 62 mg (49%) of disaccharide No. (100) are obtained according toExample B8.3(a) from 100 mg (195 μmol) of amine No. (99) and 37 mg (235μmol) of 5-fluorosalicylic acid in the presence of 33 μl (235 mmol) oftriethylamine and 89 mg (235 μmol) of HBTU instead of HBPyU in 3 ml ofdry DMF. Compound No. (100) can also be obtained analogously to ExampleB8.3(a) from amine No. (57) and 5-fluorosalicylic acid and subsequentdeacetylation with sodium methanolate, in an overall yield of 82%.

¹H-NMR (CD₃OD-CDCl₃-D₂O, 400.13 MHz) δ=1.36-1.62 (m, 8 H); 1.79-1.93 (m,4 H); 2.60 (t, 7.5 Hz, 2 H); 3.76-4.39 (m, 17 H); 4.69 (d, 8.6 Hz, 1 H);5.09 (d, 8.6 Hz, 1 H); 7.26 (dd, 4.9 Hz, 8.0 Hz, 8.3 Hz, 1 H); 7.47 (dt,3.2 Hz, 8.0 Hz, 1 H); 7.88 (dd, 3.1 Hz, 9.8 Hz, 1 H). ¹³C-NMR(CD₃OD-CDCl₃-D₂O, 100.6 MHz) δ=25.29; 26.28; 29.40; 29.50; 29.56; 29.82;34.50; 52.00; 55.88; 61.71; 61.76; 69.21; 69.56; 70.72; 71.38; 73.34;75.95; 76.13; 82.52; 101.44; 104.02; 114.06 (d, 25.7 Hz); 116.74 (d, 6.6Hz); 119.55 (d, 7.7 Hz); 121.26 (d, 23.5 Hz); 155.67 (d, 234.6 Hz);157.17; 170.00; 175.69.

(c) 20 mg (46%) of compound No. (101) are obtained according to ExampleB1.1(b) (in this case the buffer solution comprises 9% of DMSO) from 30mg (46 μmol) of compound No. (100) and 40 mg (64 μmol) of CMP-sia.

¹H-NMR (CD₃OD, 400.13 MHz) δ=0.90-1.21 (m, 8 H); 1.34-1.45 (m, 4 H);1.62 (t, 11.6 Hz, 1 H); 1.92 (s, 3 H); 2.19 (t, 7.6 Hz, 2 H); 2.71 (dd,11.6 Hz, 2.8 Hz, 1 H); 3.31 (m, 1 H); 3.37-3.71 (m, 17 H); 3.81-3.99 (m,6 H); 4.40 (d, 8.6 Hz, 1 H); 4.58 (d, 8.6 Hz, 1 H); 6.83 (dd, 5.5 Hz,10.3 Hz, 1 H); 7.08 (broad dt, 3.4 Hz, 8.1 Hz, 1 H); 7.48 (dd, 5.5 Hz,10.3 Hz, 1 H). ¹³C-NMR (CD₃OD, 100.6 MHz) δ=22.44; 25.87; 26.99; 29.93;30.14 (2×C); 30.42; 34.65; 41.63; 51.82; 53.74; 56.14; 62.39; 62.67;63.99; 68.66; 69.29; 69.53; 70.60 (2×C); 70.77; 72.58; 74.72; 76.70;77.33; 77.42; 81.87; 101.03; 102.54; 104.22; 114.53 (d, 24.5 Hz);117.50; 119.78 (d, 7.6 Hz); 121.51 (d, 23.7 Hz); 156.49 (d, 235.8 Hz);170.42; 174.96; 175.29; 175.93; no resolution of the remaining signals.

(d) 17 mg (75%) of compound No. (98) are obtained according to ExampleB2.1(c) from 20 mg (21 μmol) of compound No. (101) and 17 mg (28 μmol)of GDP-D-arabinose. ¹H-NMR (CD₃OD, 400.13 MHz) δ=0.93-1.29 (m, 8 H);1.35-1.48 (m, 4 H); 1.69 (broad t, 12.4 Hz, 1 H); 1.96 (s, 3 H); 2.19(t, 8.4 Hz, 2 H); 2.74 (dd, 12.4 Hz, 3.4 Hz, 1 H); 3.31-3.98 (m, 27 H);4.40 (broad t, 5.5 Hz, 1 H); 4.57 (m, 2 H); 4.68 (broad d, 8.6 Hz, 1 H);5.05 (d, 4.3 Hz, 1 H); 6.91 (dd, 5.5 Hz, 10.3 Hz, 1 H); 7.10 (broad dt,3.4 Hz, 8.1 Hz, 1 H); 7.48 (dd, 5.5 Hz, 10.3 Hz, 1 H). ¹³C-NMR (CD₃OD,100.6 MHz) δ=22.54; 25.98; 27.12; 30.06; 30.25; 30.26; 30.57; 34.77;41.89; 51.96; 53.91; 58.18; 61.57; 63.02; 64.01; 65.41; 70.40; 70.71(3×C); 70.98; 71.20; 74.84; 76.38 (2×C); 77.23 (2×C); 100.34; 102.24(2×C); 103.33; 115.00 (d, 24.3 Hz); 118.41; 119.93 (d, 7.6 Hz); 121.69(d, 23.2 Hz); 156.72; 156.88 (d, 234.9 Hz); 176.05; no resolution of theremaining signals. ¹⁹F-NMR (CD₃OD, 376.5 MHz) δ=−126.6.

EXAMPLE B8.7 Preparation of Compound No. (102)

(a) 42 mg (28%) of amide No. (103) are obtained according to ExampleB8.6(b) from 44 mg (258 μmol) of vanillic acid and 100 mg (234 μmol) ofcompound No. (99) in the presence of 107 mg (282 μmol) of TBTU, insteadof HBTU, and 40 μl (282 μmol) of triethylamine in 2 ml of dry DMF.Compound No. (103) can also be obtained from vanillic acid and amine No.(57) according to Example B8.1(d), in an overall yield of 27%.

¹H-NMR (CD₃OD-CDCl₃, 400.13 MHz) δ=0.99-1.60 (m, 12 H); 2.20 (t, 7.5 Hz,2 H); 3.33-3.92 (m, 19 H); 3.98 (broad t, 9.6 Hz, 1 H); 4.32 (d, 8.3 Hz,1H); 4.66 (d, 8.2 Hz, 1 H); 6.81 (d, 8.3 Hz, 1 H); 7.36 (dd, 1.2 Hz, 7.2Hz, 1H); 7.44 (d, 1.2 Hz, 1 H); ¹³C-NMR (CD₃OD-CDCl₃, 100.6 MHz)δ=25.65; 26.19; 29.53; 29.65 (2×C); 29.82; 35.38; 52.29; 56.56; 56.91;61.21 (2×C); 69.67; 69.80; 71.09; 72.13; 74.01; 74.27; 76.95; 83.35;102.12; 104.73; 112.12; 115.70; 122.06; 126.71; 148.51; 150.81; 170.63;176.30.

(b) 22 mg (70%) of compound No. (104) are obtained according to ExampleB2.1(b) (in this case the buffer solution comprises 8% of DMSO(vol/vol)) from 22 mg (34 μmol) of compound No. (103) and 58 mg (88μmol) of CMP-sia.

¹H-NMR (CD₃OD, 400.13 MHz) δ=0.93-1.29 (m, 8 H); 1.35-1.47 (m, 4 H);1.65 (broad t, 11.6 Hz, 1 H); 1.95 (s, 3 H); 2.18 (t, 7.6 Hz, 2 H); 2.73(dd, 11.6 Hz, 2.8 Hz, 1 H); 3.32 (m, 1 H); 3.37-3.71 (m, 18 H);3.80-3.98 (m, 9 H); 4.37 (d, 8.6 Hz, 1 H); 4.60 (d, 8.6 Hz, 1 H); 6.81(dd, 5.5 Hz, 10.3 Hz, 1 H); 7.33 (broad dt, 3.4 Hz, 8.1 Hz, 1 H); 7.41(dd, 5.5 Hz, 10.3 Hz, 1 H). ¹³C-NMR (CD₃OD, 100.6 MHz) δ=21.33; 24.67;25.85; 28.77; 29.03 (2×C); 29.33; 33.46; 40.43; 50.66; 52.59; 55.32(2×C); 61.38; 61.51; 62.74; 67.60; 68.11; 68.32; 69.45; 69.54 (2×C);71.45; 73.55; 75.58; 76.02; 76.22; 82.03; 99.91; 101.51; 102.98; 110.94;114.62; 120.99; 125.90; 147.44; 149.96; 169.31; 173.79; 174.13; 174.80.

(c) 10 mg (60%) of compound No. (102) are obtained according to ExampleB2.1(c) from 14 mg (15 μmol) of compound No. (104) and 14 mg (22 μmol)of GDP-D-arabinose. ¹H-NMR (CD₃OD, 400.13 MHz) δ=0.89-1.23 (m, 8 H);1.32-1.49 (m, 4 H); 1.59 (broad t, 11.0 Hz, 1 H); 1.92 (s, 3 H); 2.04(t, 7.6 Hz, 2 H); 2.70 (dd, 11.6 Hz, 2.8 Hz, 1 H); 3.30-3.93 (m, 27 H);4.30 (broad t, 9.0 Hz, 1 H); 4.48 (d, 8.6 Hz, 1 H); 4.56 (m, 2 H); 5.02(d, 4.3 Hz, 1 H); 6.81 (d, 8.3 Hz, 1 H); 7.29 (dd, 2.1 Hz, 8.3 Hz, 1 H);7.36 (d, 2.1 Hz, 1 H); ¹³C-NMR (CD₃OD, 100.6 MHz) δ=22.62; 27.19; 27.43;30.47; 30.53; 30.65; 30.70; 34.49; 42.04; 51.83; 53.88; 56.65; 58.34;61.54; 63.07; 63.86; 65.43; 68.57; 69.45 (2×C); 70.36; 70.42; 70.86;70.94; 71.14; 72.55; 74.85; 74.90; 76.53; 76.75; 77.26; 77.51; 100.40;101.13; 102.49; 103.39; 112.14; 116.25; 122.27; 126.91; 148.92; 151.48;170.63; 174.98; 175.38; no resolution of the remaining signals.

EXAMPLE B8.8 Preparation of Compound No. (105)

(a) 150 mg (293 μmol) of amine No. (99) are dissolved in 5 ml of dry DMFat RT, and 37 μl (352 μmol) of ethyl chlorothiolformate and 49 μl oftriethylamine are added in succession. After the mixture has beenstirred overnight, the solvent is evaporated off and the residue ischromatographed over silica gel (eluent: methylenechloride/methanol—4/1). 82 mg (46%) of amide No. (106) are obtained.

¹H-NMR (CD₃OD, 400.13 MHz) δ=1.15 (t, 6.7 Hz, 3 H); 1.21 (m, 8 H); 1.34(m, 4 H); 2.20 (t, 7.5 Hz, 2 H); 2.73 (broad q, 6,7 Hz, 2 H); 3.20 (m, 1H); 3.28-3.48 (m, 5 H); 3.52-3.81 (m, 11 H); 4.23 (d, 8.3 Hz, 1 H); 4.38(d, 8.2 Hz, 1 H). ¹³C-NMR (CD₃OD-CDCl₃, 100.6 MHz) 8 16.6; 24.92; 25.94;26.91; 30.00; 30.22 (2×C); 30.47; 34.81; 52.27; 57.76; 62.43; 62.48;70.08; 70.46; 70.97 72.33; 74.29; 76.95; 77.18; 83.64; 102.34; 104.77.

(b) 24 mg (69%) of compound No. (107) are obtained according to ExampleB1.1(b) (in this case the buffer solution comprises 12% of DMSO(vol/vol)) from 23 mg (39 μmol) of compound No. (106) and 35 mg (53μmol) of CMP-sia.

¹H-NMR (CD₃OD, 400.13 MHz) δ=1.20-1.36 (m, 11 H); 1.44-1.59 (m, 4 H);1.72 (broad t, 11.6 Hz, 1 H); 1.97 (s, 3 H); 2.25 (t, 7.6 Hz, 2 H);2.73-2.90 (m, 3 H); 3.23 (m, 1 H); 3.32-3.91 (m, 22 H); 3.99 (dd, 3.5Hz, 10.6 Hz, 1 H); 4.36 (d, 8.6 Hz, 1 H); 4.45 (d, 8.6 Hz, 1 H). ¹³C-NMR(CD₃OD, 100.6 MHz) δ=16.35; 22.67; 24.94; 26.04; 27.04; 30.15; 30.15;30.35; 30.37; 30.62; 34.80; 41.72; 51.98; 53.94; 57.80; 62.72 (2×C);64.21; 69.12; 69.37; 69.71; 70.50; 70.78 (2×C); 72.71; 74.91; 76.91;77.31; 77.46; 83.10; 101.24; 102.58; 104.25; 175.47 (2×C); 176.03.

(c) 14 mg (56%) of compound No. (105) are obtained according to ExampleB2.1(c) from 22 mg (25 μmol) of compound No. (107) and 20 mg (33 μmol)of GDP-D-arabinose. ¹H-NMR (CD₃OD, 400.13 MHz) δ=1.21-1.36 (m, 11 H);1.44-1.59 (m, 4 H); 1.68 (broad t, 11.0 Hz, 1 H); 1.97 (s, 3 H); 2.26(t, 7.6 Hz, 2 H); 2.71-2.94 (m, 3 H); 3.28-3.95 (m, 27 H); 4.17 (broadt, 10.0 Hz, 1 H); 4.48 (d, 7.3 Hz, 1 H); 4.57 (broad d, 7.6 Hz, 1 H);5.01 (d, 4.3 Hz, 1 H). ¹³C-NMR (CD₃OD, 100.6 MHz) δ=16.67; 22.63; 24.96;26.05; 27.05; 30.16; 30.36 (2×C); 30.65; 30.81; 42.14; 51.98; 53.95;59.66; 61.48; 63.03; 64.23; 65.34; 68.59; 69.44; 69.66; 70.35 (2×C);70.77; 70.96; 71.01; 72.50; 74.50; 74.89; 76.56; 76.70; 77.24; 77.53;100.27; 100.96; 102.01; 103.58; 170.40; 175.12; 175.44; 176.03.

EXAMPLE B8.9 Preparation of Compound No. (83)

(a) 169 mg (98%) of peracetylated disaccharide are obtained according toExample B8.12(a) from 150 mg (196 μmol) of amine No. (57) and 18 ml (236μmol) of mesyl chloride in 10 ml of methylene chloride in the presenceof 40 μl (236 μmol) of N-ethyldiisopropylamine and 2.5 mg (20 μmol) ofN,N-dimethylaminopyridine, instead of triethylamine; 52 mg (62 μmol) ofthis product are deacetylated according to Example B 8.5 with sodiummethanolte to give disaccharide No. (84). 23 mg (64%) of sulfonamide areobtained.

¹H-NMR (CD₃OD, 400.13 MHz) δ=1.14-1.29 (m, 8 H); 1.30-1.56 (m, 4 H);2.19 (t, 7.6 Hz, 2 H); 2.90 (s, 3 H); 3.02-3.85 (m, 17 H); 4.21 (d, 8.6Hz, 1 H); 4.31 (d, 8.6 Hz, 1 H). ¹³C-NMR (CD₃OD, 100.6 MHz) δ=25.97;27.13; 30.08; 30.27; 30.30; 30.70; 34.77; 42.16; 51.97; 59.86; 62.49;62.60; 70.17; 70.77; 71.00; 72.68; 74.42; 77.31; 77.36; 85.77; 102.72;105.53; 176.41.

(b) 28 mg (94%) of compound No. (85) are obtained according to ExampleB1.1(b) from 20 mg (34 μmol) of compound No. (84) and 49 mg (55 μmol) ofCMP-sia.

¹H-NMR (CD₃OD, 400.13 MHz) δ=1.21-1.38 (m, 8 H); 1.49-1.61 (m, 4 H);1.72 (broad t, 12.4 Hz, 1 H); 1.96 (s, 3 H); 2.26 (t, 7.6 Hz, 2 H); 2.79(broad dd, 12.4 Hz, 3.4 Hz, 1H)3.01 (s, 3 H); 3.13-3.95 (m, 24 H ); 4.28(d, 8.6 Hz, 1 H); 4.40 (broad d, 8.6 Hz, 1 H). ¹³C-NMR (CD₃OD, 100.6MHz) δ=22.72; 25.98; 27.05; 30.08; 30.26; 30.30; 30.66; 34.78; 41.84;42.47; 51.97; 53.90; 59.77; 62.61; 62.74; 64.30; 69.03; 69.39; 69.95;70.79; 71.00; 71.25; 71.30; 72.94; 74.81; 77.15; 77.36; 84.37; 100.18;103.01; 104.89; 175.41; 176.03 (2×C).

(c) 12 mg (67%) of compound No. (83) are obtained according to ExampleB2.1(c) from 15 mg (17 μmol) of compound No. (85) and 17 mg (28 μmol) ofGDP-D-arabinose. ¹H-NMR (CD₃OD, 400.13 MHz) δ=1.18-1.32 (m, 8 H);1.40-1.58 (m, 4 H); 1.65 (t, 12.4 Hz, 1 H); 1.95 (s, 3 H); 2.24 (t, 8.4Hz, 2 H); 2.82 (dd, 12.4 Hz, 3.4 Hz, 1 H); 3.01 (s, 3 H); 3.28-3.95 (m,27 H); 3.99 (dd, 3.4 Hz, 8.3 Hz, 1 H); 4.29 (d, 8.6 Hz, 1 H); 4.63(broad d, 8.6 Hz, 1 H); 5.02 (d, 4.8 Hz, 1 H); the remaining signals aremasked by the solvent. ¹³C-NMR (CD₃OD, 100.6 MHz) δ=22.63; 25.98; 27.10;30.08; 30.27; 30.31; 30.68; 34.78; 42.46; 43.06; 51.99; 53.92; 61.24;62.15; 63.10; 64.41; 65.49; 68.55; 69.39; 69.91; 70.24; 70.36; 70.97;71.05; 71.41; 72.42; 73.64; 74.45; 74.83; 76.91; 77.27; 77.59; 100.42;100.79; 102.90; 103.94; 174.80; 175.45; 176.04.

EXAMPLE B8.10 Preparation of Compound No. (89)

14 mg (84%) of compound No. (89) are obtained according to ExampleB2.1(c) from 14 mg (15 μmol) of compound No. (85) and 17 mg (28 μmol) ofGDP-L-galactose. ¹H-NMR (CD₃OD, 400.13 MHz) δ=0.85-1.18 (m, 8 H);1.34-1.47 (m, 4 H); 1.6 (t, 12.4 Hz, 1 H); 1.96 (s, 3 H); 2.20 (t, 8.4Hz, 2 H); 2.26 (s, 3 H); 2.68 (dd, 12.4 Hz, 3.4 Hz, 1 H); 3.38 (m, 1 H);3.41-4.00 (m, 27 H); 4.39 (broad t, 7.0 Hz, 1 H); 5.09 (d, 4.3 Hz, 1 H);6.84 (d, 7.6 Hz, 1 H); 7.20 (dd, 1.4 Hz, 7.6 Hz, 1 H); 7.58 (d, 1.4 Hz,1 H); the remaining signals are masked by the solvent. ¹³C-NMR (CD₃OD,100.6 MHz) δ=20.73; 22.81; 25.75; 26.82; 29.75; 29.86; 29.96; 30.21;34.76; 41.08; 52.49; 53.44; 61.31; 61.90; 62.63; 63.95; 68.81; 69.20;69.31; 69.85; 70.51; 70.60; 70.66; 71.00; 71.10; 72.51; 74.25; 74.42;75.81; 76.15; 76.88 (2×C); 77.24; 99.51; 101.16; 102.18; 103.65; 117.31;118.38; 129.38; 129.92; 135.89; 157.72; 171.36; 175.10; 175.88; 177.13.

EXAMPLE B8.11 Preparation of Compound No. (61)

(a) 104 mg (63%) of amide No. (62) are obtained according to ExampleB8.1(d) from 45 mg (288 μmol) of orotic acid and 200 mg (262 μmol) ofcompound No. (57).

¹H-NMR (D₆-DMSO, 250.13 MHz) δ=1.18 (m, 8 H); 1.43 (m, 4 H); 2.26 (t,7.5 Hz, 2 H); 3.17-3.79 (m, 17 H); 4.14 (m, 3 H); 4.48 (d, 8.6 Hz, 1 H);4.55 (d, 8.6 Hz, 1 H); 4.67 (m, 2 H); 4.83 (s, 1 H); 4.88 (m, 1 H); 6.00(s, 1 H); 8.75 (broad, 1 H). ¹³C-NMR (D₆-DMSO, 62.89 MHz) δ=24.51;25.57; 28.52; 28.78; 28.86; 29.09; 33.34; 48.67; 54.48; 60.50 (2×C);68.71; 68.76; 69.32; 70.40; 73.20; 75.70; 76.53; 84.49; 99.47; 100.39;104.40; 146.61; 151.84; 160.96; 164.46; 173.50.

(b) 55 mg (73%) of compound No. (63) are obtained according to ExampleB1.1(b) (in this case the reaction mixture comprises 8% of DMSO(vol/vol)) from 50 mg (77 μmol) of compound No. (62) and 69 mg (104μmol) of CMP-sia.

¹H-NMR (CD₃OD, 250.13 MHz) δ=1.18 (m, 8 H); 1.46 (m, 4 H); 1.66 (broadt, 11.0 Hz, 1 H); 1.93 (s, 3 H); 2.22 (t, 7.6 Hz, 2 H); 2.73 (broad d,11.0 Hz, 1 H); 3.26-4.00 (m, 24 H); 4.31 (d, 8.6 Hz, 1 H); 4.52 (d, 8.6Hz, 1 H); 6.10 (s, 1 H). ¹³C-NMR (CD₃OD, 62.89 MHz) δ=22.78; 26.00;27.91; 30.10; 30.32; 30.44; 30.59; 34.77; 41.62; 52.02; 53.89; 56.57;62.64 (2×C); 64.29; 69.01; 69.28; 69.84; 70.77 (3×C); 73.10; 74.86;76.79; 77.36; 77.51; 84.17; 101.13 (2×C); 102.14; 105.35; 148.07;153.97; 162.74; 167.10; 175.15; 175.46; 176.11.

(c) 11 mg (84%) of compound No. (61) are obtained according to ExampleB2.1(c) from 12 mg (12.0 μmol) of compound No. (63) and 10 mg (14 μmol)of GDP-D-arabinose. ¹H-NMR (CD₃OD, 400.13 MHz) δ=1.22 (m, 8 H); 1.48 (m,4 H); 1.73 (t, 11.0 Hz, 1 H); 1.96 (s, 3 H); 2.25 (t, 7.6 Hz, 2 H); 2.75(dd, 11.0 Hz, 3.4 Hz, 1 H); 3.33-3.94 (m, 27 H); 4.21 (t, 9.9 Hz, 1 H);4.44 (d, 8.6 Hz, 2 H); 4.57 (m, 2 H); 5.06 (d, 4.8 Hz, 1 H); 6.10 (s, 1H). ¹³C-NMR (CD₃OD, 100.60 MHz) δ=22.60; 26.00; 27.25; 30.13; 30.34;30.44; 30.64; 34.79; 41.61; 51.97; 53.97; 58.29; 61.55; 63.15; 64.48;65.40; 69.06; 69.66; 70.03; 70.35 (2×C); 70.76; 70.97; 71.06; 72.97;74.44; 74.95; 76.57; 77.03; 77.38; 77.50; 100.23; 100.63; 102.09;103.85; 104.06; no resolution of the remaining signals.

EXAMPLE B8.12 Preparation of Compound No. (74)

(a) 250 mg (327 μmol) of amine No. (57) are dissolved in 5 ml ofmethylene chloride at RT to give a clear solution, and 57 μl (490 μmol)of benzoyl chloride and 78 μl of triethylamine are added. After themixture has been stirred overnight, the solvent is evaporated off andthe residue is chromatographed over silica gel (eluent: ethylacetate/hexane—6/4). 197 mg (70%) of peracetylated disaccharide areobtained and are dissolved in 2 ml of absolute methanol and deacetylatedwith 0.5 ml of a 0.5% sodium methanolate solution. After 1.5 h, thereaction mixture is evaporated and the residue is chromatographed oversilica gel (eluent: methylene chloride/methanol/water—10/4/0.8). 76 mg(57%) of disaccharide No. (75) are obtained.

¹H-NMR (CD₃D-CDCl₃-D₂O, 400.13 MHz) δ=1.05 (very broad m, 8 H); 1.40 (m,4 H); 2.18 (t, 7.6 Hz, 2 H); 3.31-3.40 (m, 2 H); 3.40-3.51 (m, 4 H);3.59 (s, 3 H); 3.60 (dd, 4.4 Hz, 12.1 Hz, 1 H); 3.62-3.73 (m, 4 H); 4.26(d, 8.6 Hz, 1 H); 4.66 (broad d, 8.6 Hz, 1 H); 7.39 (t, 8.8 Hz, 2 H);7.46 (broad t, 8.8 Hz, 1 H); 7.76 (broad d, 8.8 Hz, 2 H). ¹³C-NMR(CD₃OD-CDCl₃-D₂O, 100.61 MHz) δ=25.65; 26.73; 29.70; 29.86; 29.92;30.24; 34.72; 52.25; 56.66; 62.12; 62.27; 66.69; 70.23; 70.88; 71.90;73.89; 76.53; 76.82; 83.40; 101.93; 104.71; 128.25 (2×C); 129.35 (2×C);132.60; 135.26; 170.95; 176.31.

(b) 33 mg (72%) of compound No. (76) are obtained according to ExampleB1.1(b) from 31 mg (51 μmol) of compound No. (75) and 46 mg (69 μmol) ofCMP-sia.

¹H-NMR (CD₃OD, 400.13 MHz) δ=1.10 (broad m, 8 H); 1.41 (m, 4 H); 1.64(broad t, 11.6 Hz, 1 H); 1.93 (s, 3 H); 2.17 (t, 7.6 Hz, 2 H); 2.71 (dd,11.6 Hz, 2.8 Hz, 1 H); 3.30-3.73 (m, 18 H); 3.80-3.94 (m, 6 H); 4.35 (d,8.6 Hz, 1 H); 4.60 (broad d, 8.6 Hz, 1 H); 7.42 (m, 3 H); 7.77 (m, 1 H).¹³C-NMR (CD₃OD, 100.61 MHz) δ=22.72; 25.87; 27.12; 30.05; 30.26; 30.28;30.62; 34.76; 41.39; 51.97; 53.92; 56.65; 62.59; 62.79; 64.04; 69.27(2×C); 69.66; 70.74 (3×C); 72.74; 74.84; 76.68; 77.29; 77.50; 83.83;101.36; 102.67; 104.59; 128.58 (2×C); 129.61 (2×C); 132.58; 136.23;166.15; 175.29; 175.48; 175.99.

(c) 18 mg (87%) of compound No. (74) are obtained according to ExampleB2.1(c) from 18 mg (19 μmol) of compound No. (76) and 18 mg (27 μmol) ofGDP-L-galactose. ¹H-NMR (CD₃OD, 400.13 MHz) δ=1.05 (broad m, 8 H); 1.40(m, 4 H); 1.62 (broad t, 12.4 Hz, 1 H); 1.94 (s, 3 H); 2.16 (t, 8.4 Hz,2 H); 2.68 (dd, 12.4 Hz, 3.4 Hz, 1 H); 3.29 (broad t, 11.0 Hz, 1 H);3.37-3.96 (m, 27 H); 4.31(broad t, 15.4 Hz, 1 H); 4.47 (d, 8.6 Hz, 1 H);4.63 (broad t, 11.0 Hz, 1 H); 4.72 (broad d, 8.6 Hz, 1 H); 5.05 (d, 4.8Hz, 1 H); 7.46 (m, 3 H); 7.77 (m, 2 H). ¹³C-NMR (CD₃OD, 100.61 MHz)δ=22.78; 25.80; 26.92; 29.80; 29.89; 30.05; 30.38; 34.76; 41.40; 52.39;53.56; 62.00 (2×C); 62.69; 63.82; 68.96; 69.40; 69.99; 70.63; 70.73;70.94; 71.03 (3×C); 72.50; 74.35; 74.56; 76.22; 76.98; 77.17; 99.63;101.43; 104.01; 128.41 (2×C); 129.92 (2×C); 133.06; 135.32; 175.05;175.89; 176.91; no resolution of the remaining signals.

EXAMPLE B8.13 Preparation of Compound No. (86)

(a) 119 mg (57%) of peracetylated amide are obtained according toExample A8 from 180 mg (236 μmol) of amine No. (57) and 39 mg (256 μmol)of 5-methylsalicylic acid in 3 ml of dry acetonitrile, and the productis deacetylated immediately according to Example B8.1(d): 77 mg (90%) ofdisaccharide No. (87) are obtained.

¹H-NMR (CD₃OD-CDCl₃, 400.13 MHz) δ=0.90-1.21 (m, 8 H); 1.33-1.54 (m, 4H); 2.19 (t, 7.5 Hz, 2 H); 2.21 (s, 3 H); 3.32-3.99 (m, 17 H); 4.32 (d,8.6 Hz, 1 H); 4.64 (d, 8.6 Hz, 1 H); 7.78 (d, 7.6 Hz, 1 H); 8.15 (dd,1.4 Hz, 7.6 Hz, 1 H); 8.60 (d, 1.4 Hz, 1 H). ¹³C-NMR (CD₃OD-CDCl₃, 100.6MHz) δ=20.68; 25.57; 26.59; 29.65; 29.73; 29.86; 30.12; 34.69; 52.14;56.01; 62.03; 62.18; 69.59; 70.06; 70.92; 71.80; 73.71; 76.32; 76.62;83.19; 102.07; 104.37; 116.77; 119.15; 127.54; 128.89; 135.26; 160.61;171.70; 176.11.

(b) 31 mg (84%) of compound No. (88) are obtained according to ExampleB1.1(b) (in this case the buffer solution contains 9% of DMSO (vol/vol))from 25 mg (39 μmol) of compound No. (87) and 35 mg (53 μmol) ofCMP-sia.

¹H-NMR (CD₃OD, 400.13 MHz) δ=0.89-1.21 (m, 8 H); 1.34-1.46 (m, 4 H);1.65 (t, 11.6 Hz, 1 H); 1.96 (s, 3 H); 2.18 (t, 7.6 Hz, 2 H); 2.28 (s, 3H); 2.72 (dd, 11.6 Hz, 2.8 Hz, 1 H); 3.33 (m, 1 H); 3.39-3.74 (m, 17 H);3.81-4.02 (m, 6 H); 4.41 (d, 8.6 Hz, 1 H); 4.61 (broad d, 8.6 Hz, 1 H);6.75 (d, 7.6 Hz, 1 H); 7.16 (dd, 1.4 Hz, 7.6 Hz, 1 H); 7.54 (d, 1.4 Hz,1 H). ¹³C-NMR (CD₃OD, 100.6 MHz) δ=20.57; 22.46; 25.80; 26.92; 29.87;30.02; 30.09 (2×C); 30.35; 34.58; 41.37; 51.76; 53.70; 55.95; 62.26;62.57; 63.73; 68.68; 69.10; 69.37; 70.49; 70.57; 70.65; 72.51; 74.64;76.52; 77.11; 77.28; 82.48; 101.03; 101.52; 103.92; 116.46; 118.25;128.64; 129.05; 135.46; 159.08; 171.69; 174.96; 175.26; 175.83.

(c) 14 mg (84%) of compound No. (86) are obtained according to ExampleB2.1(c) from 14 mg (15 μmol) of compound No. (88) and 16 mg (26 μmol) ofGDP-D-arabinose. ¹H-NMR (CD₃OD, 400.13 MHz) δ=0.91-1.20 (m, 8 H);1.34-1.46 (m, 4 H); 1.62 (broad t, 12.4 Hz, 1 H); 1.96 (s, 3 H); 2.19(t, 8.4 Hz, 2 H); 2.26 (s, 3 H); 2.75 (broad dd, 12.4 Hz, 3.4 Hz, 1 H);3.29 (m, 1 H); 3.36-4.00 (m, 27 H); 4.42 (broad t, 7.0 Hz, 1 H); 4.58(broad d, 8.6 Hz, 1 H); 4.68 (broad d, 8.6 Hz, 1 H); 5.08 (d, 4.3 Hz, 1H); 6.81 (d, 7.6 Hz, 1 H); 7.17 (dd, 1.4 Hz, 7.6 Hz, 1 H); 7.53 (d, 1.4Hz, 1 H). ¹³C-NMR (CD₃OD, 100.6 MHz) δ=20.77; 22.63; 26.00; 27.13;30.08; 30.20; 30.28; 30.59; 34.79; 41.94; 51.96; 53.93; 57.94; 61.59;63.03; 63.93; 65.41; 68.67; 69.36; 69.50; 70.37; 70.44; 70.74; 70.92;71.14; 72.53; 74.87; 74.96; 76.47 (2×C); 77.22; 77.51; 100.39; 101.15;102.38; 103.21; 117.13; 118.58; 129.03; 129.40; 135.71; 158.84; 171.67;175.05; 175.45; 176.04.

EXAMPLE B8.14 Preparation of Compound No. (90)

(a) 150 mg (196 μmol) of amine No. (57) are dissolved in 2 ml of drymethylene chloride at RT, and 70 μl (503 μmol) of trifluoroaceticanhydride and 70 μl of triethylamine are added in succession. After 3 h,the solvent is evaporated off and the residue is chromatographed oversilica gel (eluent: hexane/ethyl acetate—1/1). 156 mg (93%) ofperacetylated amide are obtained and are deacetylated by means of sodiummethanolate—as described in Example 30(a). 110 mg (100%) of disaccharideNo. (91) are obtained.

¹H-NMR (CD₃OD-CDCl₃, 400.13 MHz) δ=1.21-1.38 (m, 8 H); 1.46-1.62 (m, 4H); 2.30 (t, 7.5 Hz, 2 H); 3.33 (m, 1 H); 3.42-3.92 (m, 16 H); 4.38 (d,8.6 Hz, 1 H); 4.53 (broad d, 8.6 Hz, 1 H). ¹³C-NMR (CD₃OD-CDCl₃, 100.6MHz) δ=25.84; 26.78; 29.90; 30.06; 30.09; 30.37; 34.74; 52.09; 56.49;62.28; 62.38; 69.92; 70.31; 70.77; 72.00; 74.45; 76.89; 77.27; 83.40;101.62; 105.09; 176.24; no resolution of the remaining signals.

(b) 39 mg (89%) of compound No. (92) are obtained according to ExampleB1.1(b) (in this case the buffer solution comprises 9% of DMSO(vol/vol)) from 30 mg (49 μmol) of compound No. (91) and 42 mg (64 μmol)of CMP-sia.

¹H-NMR (CD₃OD, 400.13 MHz) δ=1.19-1.32 (m, 8 H); 1.41-1.56 (m, 4 H);1.71 (t, 11.6 Hz, 1 H); 1.97 (s, 3 H); 2.25 (t, 7.6 Hz, 2 H); 2.74 (dd,11.6 Hz, 2.8 Hz, 1 H); 3.28 (m, 1 H); 3.35-3.97 (m, 23 H); 4.31 (d, 8.6Hz, 1 H); 4.45 (broad d, 8.6 Hz, 1 H). ¹³C-NMR (CD₃OD, 100.6 MHz)δ=22.73; 25.99; 26.96; 30.06; 30.24 (2×C); 30.56; 34.77; 41.43; 51.97;53.97; 56.37; 63.51; 62.61; 63.81; 69.26; 69.53; 70.48 (2×C); 70.54;70.72; 72.69; 74.87; 76.71; 77.58 (2×C); 83.45; 101.33; 101.95; 104.82;117.51 (q); 159.40 (q); 175.32; 175.50; 176.05.

(c) 16 mg (84%) of compound No. (90) are obtained according to ExampleB2.1(c) from 17 mg (19 μmol) of compound No. (92) and 18 mg (29 μmol) ofGDP-D-arabinose. ¹H-NMR (CD₃OD, 400.13 MHz) δ=1.14-1.28 (m, 8 H);1.40-1.55 (m, 4 H); 1.65 (t, 12.4 Hz, 1 H); 1.95 (s, 3 H); 2.25 (t, 8.4Hz, 2 H); 2.74 (dd, 12.4 Hz, 3.4 Hz, 1 H); 3.30-3.90 (m, 28 H); 4.10 (t,7.0 Hz, 1 H); 4.37 (d, 8.6 Hz, 1 H); 4.48 (d, 8.6 Hz, 1 H); 5.01 (d, 4.3Hz, 1 H). ¹³C-NMR (CD₃OD, 100.6 MHz) δ=22.77; 25.87; 26.79; 29.87;30.03; 30.06; 30.37; 32.51; 41.90; 52.35; 53.70; 57.84; 61.18; 62.95;63.47; 65.27; 68.19; 69.14; 69.40; 69.94; 70.06; 70.59; 70.79; 70.99;72.34; 73.97; 74.60; 76.37; 76.54; 77.14; 77.45; 100.12; 100.72; 101.70;103.97; 117.39 (q); 159.41 (q); 175.02; 175.77; 176.85.

EXAMPLE B9.1 Preparation of Compound No. (27)

(a) 19 mg (95%) of compound No. (29) are obtained according to ExampleB1.1 (a1) (in this case the buffer solution comprises about 11% of DMSO(vol/vol)) from 15 mg (31 μmol) of compound No. (28) and 25 mg (40 μmol)of UDP-gal.

¹H-NMR (CD₃OD, 250.13 MHz) δ=1.01 (m, 8 H); 1.39 (m, 4 H); 2.13 (t, 7.5Hz, 2 H); 2.21 (s, 3 H); 3.32-3.95 (m, 17 H); 4.32 (d, 8.6 Hz, 1 H);4.50 (d, 8.6 Hz, 1 H); 6.71 (d, 7.6 Hz, 1 H); 7.11 (dd, 1.4 Hz, 7.6 Hz,1 H); 7.52 (d, 1.4 Hz, 1 H). ¹³C-NMR (CD₃OD, 62.90 MHz) δ=20.60; 26.00;27.12; 30.07; 30.28 (2×C); 30.56; 34.77; 51.95; 56.64; 62.00; 62.57;70.36; 70.75; 72.63; 73.92; 74.83; 76.59; 77.18; 81.15; 102.83; 105.11;116.37; 118.44; 128.36; 129.05; 135.59; 171.77; 176.03.

(b) 26 mg (99%) of compound No. (30) are obtained according to ExampleB1.1(b) (in this case the buffer solution comprises 8% of DMSO(vol/vol)) from 18 mg (28 μmol) of compound No. (29 and 28 mg (43 μmol)of CMP-sia.

¹H-NMR (CD₃OD, 250.13 MHz) δ=1.02 (m, 8 H); 1.38 (m, 4 H); 1.66 (broadt, 11.6 Hz, 1 H); 1.94 (s, 3 H); 2.14 (t, 7.6 Hz, 2 H); 2.19 (s, 3 H);2.78 (dd, 11.6 Hz, 2.8 Hz, 1 H); 3.32-4.01 (m, 24 H); 4.41 (d, 8.6 Hz, 1H); 4.49 (d, 8.6 Hz, 1 H); 6.66 (d, 7.6 Hz, 1 H); 7.06 (dd, 1.4 Hz, 7.6Hz, 1 H); 7.53 (d, 1.4 Hz, 1 H). ¹³C-NMR (CD₃OD, 62.90 MHz) δ=20.61;22.58; 26.01; 27.11; 30.08; 30.26; 30.31; 30.59; 34.79; 42.10; 51.95;53.93; 56.61; 62.01; 62.79; 64.54; 69.05; 69.34; 70.07; 70.84; 72.97;74.23; 74.93 (2×C); 76.52; 77.12; 77.63; 81.11; 101.06; 103.09; 104.96;117.07; 119.46; 127.56; 128.84; 135.34; 162.28; 171.99; 174.91; 175.49;176.05.

(c) 6 mg (54%) of compound No. (27) are obtained according to ExampleB3.1(c) from 11 mg (12 μmol) of compound No. (30) and 11 mg (19 μmol) ofGDP-D-arabinose. ¹H-NMR (CD₃OD, 250.13 MHz) δ=1.02 (m, 8 H); 1.48 (m, 4H); 1.62 (broad t, 11.0 Hz, 1 H); 1.93 (s, 3 H); 2.14 (t, 7.6 Hz, 2 H);2.20 (s, 3 H); 2.79 (dd, 11.6 Hz, 2.8 Hz, 1 H); 3.62-4.17 (m, 28 H);4.46 (d, 8.6 Hz, 1 H); 4.53 (m, 2 H); 5.06 (d, 4.3 Hz, 1 H); 6.70 (d,7.6 Hz, 1 H); 7.12 (dd, 1.4 Hz, 7.6 Hz, 1 H); 7.49 (d, 1.4 Hz, 1 H).¹³C-NMR (CD₃OD, 62.90 MHz) δ=20.61; 22.57; 26.00; 27.14; 30.07; 30.22;30.29; 30.58; 34.79; 42.35; 51.95; 53.96; 57.67; 61.38; 62.97; 64.64;65.12; 68.86; 69.29; 70.14 (3×C); 70.76; 70.97 (2×C); 73.03; 75.02;75.32; 75.67; 76.84; 77.29; 77.90; 99.89; 100.86; 102.62; 103.77;116.59; 118.97; 128.48; 129.19; 135.68; 159.56; 171.67; 174.84; 175.51;176.04.

EXAMPLE B9.2 Preparation of Compound No. (31)

14 mg (82%) of compound No. (31) are obtained according to ExampleB3.1(c) from 15 mg (16 μmol) of compound No. (30) and 17 mg (26 μmol) ofGDP-L-galactose. ¹H-NMR (CD₃OD, 400.13 MHz) δ=0.50 (m, 2 H); 1.06 (m, 4H); 1.16 (m, 2 H); 1.42 (m, 4 H); 1.77 (broad t, 11.0 Hz, 1 H); 2.07 (s,3 H); 2.18 (t, 7.6 Hz, 2 H); 2.24 (s, 3 H); 2.84 (dd, 11.6 Hz, 2.8 Hz, 1H); 3.37-3.73 (m, 17 H); 3.80-3.85 (m, 8 H); 4.06 (m, 4 H); 4.53 (d, 8.6Hz, 1 H); 4.57 (broad d, 8.4 Hz, 1 H); 4.70 (t, 6.4 Hz, 1 H); 5.08 (d,4.3 Hz, 1 H); 6.74 (d, 7.6 Hz, 1 H); 7.16 (dd, 1.4 Hz, 7.6 Hz, 1 H);7.54 (d, 1.4 Hz, 1 H). ¹³C-NMR (CD₃OD, 100.60 MHz) δ=20.62; 22.60;26.00; 27.13; 30.07; 30.20; 30.27; 30.57; 34.79; 42.28; 51.95; 53.98;57.43; 61.24; 62.41; 62.68; 64.63; 68.14; 69.06; 69.27; 70.12 (2×C);70.76; 70.92; 70.97; 71.09; 73.05; 75.02; 75.84; 76.39; 76.75; 77.35;77.68; 100.00; 100.93; 102.53; 104.07; 116.63; 118.51; 128.51; 129.22;135.69; 159.63; 171.81; 174.84; 175.52 (2×C).

EXAMPLE B10.1 Preparation of Compound No. (32)

(a) 27 mg (83%) of compound No. (34) are obtained according to ExampleB1.1 (a1) (in this case the buffer solution comprises about 8% of DMSO(vol/vol)) from 29 mg (54 μmol) of compound No. (33) and 39 mg (63 μmol)of UDP-gal.

¹H-NMR (CD₃OD, 400.13 MHz) δ=1.19 (m, 6 H); 1.22 (m, 2 H); 1.42 (m, 4H); 2.17 (t, 7.5 Hz, 2 H); 3.33-3.89 (m, 17 H); 4.33 (d, 8.6 Hz, 1 H);4.49 (d, 9.0 Hz, 1 H); 6.35 (t, about 2.0 Hz, 1 H); 6.66 (d, about 2.0Hz, 2 H). ¹³C-NMR (CD₃OD, 100.61 MHz) δ=26.00; 27.16; 30.10; 30.30;30.35; 30.67; 34.79; 51.96; 57.20; 60.06; 62.54; 70.34; 70.76; 72.63;73.92; 74.84; 76.56; 77.15; 81.29; 102.83; 105.12; 106.43; 106.90 (2×C);138.31; 159.73 (2×C); 170.84; 176.21.

(b) 27 mg (68%) of compound No. (35) are obtained according to ExampleB1.1(b) from 27 mg (42 μmol) of compound No. (34) and 39 mg (59 μmol) ofCMP-sia.

¹H-NMR (CD₃OD, 250.13 MHz) δ=1.08 (m, 8 H); 1.48 (m, 4 H); 1.63 (broadt, 11.0 Hz, 1 H); 1.90 (s, 3 H); 2.12 (t, 7.6 Hz, 2 H); 2.73 (dd, 11.0Hz, 2.8 Hz, 1 H); 3.38-3.88 (m, 23 H); 3.95 (dd, 10.0 Hz, 3.4 Hz, 1 H);4.35 (d, 8.6 Hz, 1 H); 4.41 (d, 8.6 Hz, 1 H); 6.29 (t, about 2.0 Hz, 1H); 6.65 (d, about 2.0 Hz, 2 H). ¹³C-NMR (CD₃OD, 100.61 MHz) δ=22.65;26.00; 27.15; 30.10; 30.30; 30.36; 30.66; 34.79; 41.58; 51.96; 53.94;57.00; 62.05; 62.75; 64.42; 69.12; 69.29; 70.01; 70.79; 70.87; 72.96;73.97; 74.90; 76.51; 77.12; 77.61; 81.37; 101.11; 102.91; 105.00;106.46; 106.92 (2×C); 138.26; 159.74 (2×C); 170.87; 175.03; 175.50;176.22.

(c) 11 mg (87%) of compound No. (32) are obtained according to ExampleB3.1(c) from 11 mg (12 μmol) of compound No. (35) and 12 mg (20 μmol) ofGDP-D-arabinose. ¹H-NMR (CD₃OD, 250.13 MHz) δ=1.09 (m, 8 H); 1.41 (m, 4H); 1.62 (broad t, 11.0 Hz, 1 H); 1.92 (s, 3 H); 2.18 (t, 7.6 Hz, 2 H);2.79 (dd, 2.8 Hz, 11.0 Hz, 1 H); 3.27-4.06 (m, 28 H); 4.43-4.60 (m, 3H); 5.08 (d, 5.0 Hz, 1 H); 6.32 (t, about 3.0 Hz, 1 H); 6.65 (d, about 3Hz, 2 H). ¹³C-NMR (CD₃OD, 62.90 MHz) δ=22.58; 26.01; 27.20; 30.12;30.28; 30.38; 30.69; 34.81; 42.81; 51.96; 53.96; 58.23; 61.35; 62.97;64.64; 65.48; 68.84; 69.29; 70.18 (2×C); 70.78; 70.96 (2×C); 73.03;75.07; 75.39; 76.27; 76.80; 77.25; 77.80; 99.89; 100.85; 102.52; 103.81;106.61; 106.94 (2×C); 138.05; 159.85 (2×C); 170.92; 175.51; noresolution of the remaining signals.

EXAMPLE B10.2 Preparation of Compound No. (36)

9 mg (70%) of compound No. (36) are obtained according to ExampleB3.1(c) from 11 mg (12 μmol) of compound No. (35) and 12 mg (18 μmol) ofGDP-L-galactose. ¹H-NMR (CD₃OD, 250.13 MHz) δ=1.09 (m, 8 H); 1.46 (m, 4H); 1.62 (broad t, 11.0 Hz, 1 H); 1.92 (s, 3 H); 2.18 (t, 7.6 Hz, 2 H);2.80 (dd, 2.8 Hz, 11.0 Hz, 1 H); 3.30-4.08 (m, 29 H); 4.48 (d, 8.6 Hz, 1H); 4.50 (broad d, 8.6 Hz, 1 H); 4.65 (t, 6.4 Hz, 1 H); 5.04 (d, 5.0 Hz,1 H); 6.32 (t, about 3.0 Hz, 1 H); 6.65 (d, about 3 Hz, 2 H). ¹³C-NMR(CD₃OD, 62.90 MHz) δ=22.58; 26.01; 27.20; 30.12; 30.29; 30.38; 30.69;34.81; 42.23; 52.15; 53.96; 58.44; 61.21; 62.43; 62.70; 64.66; 69.00;69.27; 70.15 (2×C); 70.79; 70.92 (2×C); 71.05 (2×C); 73.05; 75.02;75.87; 76.31; 76.78; 77.31; 77.66; 99.90; 100.90; 102.44; 104.09;106.62; 106.95 (2×C); 138.04; 159.85 (2×C); 171.03; 174.81; 175.52;176.23.

EXAMPLE B11 Preparation of Compound No. (37)

(a) 13 mg (41%) of compound No. (38) are obtained according to ExampleB1.1(a1) (in this case the buffer solution comprises about 9% of DMSO(vol/vol)) from 22 mg (46 μmol) of compound No. (37a) and 36 mg (59μmol) of UDP-gal.

¹H-NMR (CD₃OD, 250.13 MHz) δ=1.15 (m, 8 H); 1.51 (m, 4 H); 2.27 (t, 7.5Hz, 2 H); 3.41-4.02 (m, 17 H); 4.43 (d, 8.6 Hz, 1 H); 4.62 (d, 8.6 Hz, 1H); 6.92 (dd, 5.5 Hz, 10.3 Hz, 1 H); 7.16 (ddd, 3.4 Hz, 7.6 Hz, 8.3 Hz,1 H); 7.59 (dd, 5.5 Hz, 10.3 Hz, 1 H). ¹³C-NMR (CD₃OD, 62.90 MHz)δ=25.45; 26.46; 29.54; 29.67;29.71; 29.98; 34.59; 52.06; 56.19; 61.42;61.96; 69.59; 70.72; 71.98; 73.05; 74.00; 75.69; 76.35; 80.47; 102.00;104.23; 113.94 (d, 24.7 Hz); 117.11 (d, 6.5 Hz); 119.38 (d, 7.4 Hz);121.38 (d, 23.3 Hz); 156.97 (d, 174.2 Hz); 158.63; 170.20; 175.96.

(b) 11 mg (65%) of compound No. (39) are obtained according to ExampleB1.1(b) (in this case the buffer solution comprises 8% of DMSO(vol/vol)) from 12 mg (18 μmol) of compound No. (38) and 22 mg (33 μmol)of CMP-sia.

¹H-NMR (CD₃OD, 400.13 MHz) δ=1.00 (m, 8 H); 1.32 (m, 4 H); 1.62 (broadt, 11.6 Hz, 1H); 1.89 (s, 3 H); 2.09 (t, 7.6 Hz, 2 H); 2.71 (dd, 11.6Hz, 2.8 Hz, 1 H); 3.29-3.95 (m, 24 H); 4.35 (d, 8.6 Hz, 1 H); 4.42 (d,8.6 Hz, 1 H); 6.76 (dd, 5.5 Hz, 10.3 Hz, 1 H); 7.03 (ddd, 3.4 Hz, 7.6Hz, 8.3 Hz, 1 H); 7.45 (dd, 5.5 Hz, 10.3 hz, 1 H). ¹³C-NMR (CD₃OD, 62.90MHz) δ=22.63; 26.00, 27.13; 30.07; 30.31 (2×C); 30.55; 34.77; 42.15;51.96; 53.98; 56.68; 62.35; 62.77;, 64.36; 69.28 (2×C); 70.01; 70.76;70.88; 72.98; 73.93; 74.93; 76.57; 76.98; 77.64; 81.23; 101.21; 102.81;104.99; 113.96 (d, 24.7 Hz); 119.87 (d, 7.4 Hz); 121.56 (d, 23.3 Hz);156.55 (d, 174.2 Hz); 175.18; 175.51; no resolution of the remainingsignals.

(c) 8 mg (46%) of compound No. (37) are obtained according to ExampleB3.1(c) from 15 mg (16 μmol) of compound No. (39) and 16 mg (26 μmol) ofGDP-D-arabinose. ¹H-NMR (CD₃OD, 400.13 MHz) δ=1.07 (m, 8 H); 1.43 (m, 4H); 1.67 (broad t, 11.0 Hz, 1 H); 1.96 (s, 3 H); 2.18 (t,7.6Hz, 2 H);2.83 (dd, 11.6 Hz, 2.8 Hz, 1 H); 3.32-4.16 (m, 28 H); 4.51 (d, 8.6 Hz, 1H); 4.55 (t, 6.4 Hz, 1 H); 4.59 (d, 8.6 Hz, 1 H); 5.08 (d, 4.3 Hz, 1 H);6.85 (dd, 5.5 Hz, 10.3 Hz, 1 H); 7.13 (ddd, 3.4 Hz, 7.6 Hz, 8.3 Hz, 1H); 7.49 (dd, 5.5 Hz, 10.3 Hz, 1 H). ¹³C-NMR (CD₃OD, 100.60 MHz)δ=22.57; 26.99; 27.13; 30.05; 30.25 (2×C); 30.57; 34.78; 42.37; 51.95;53.97; 57.45; 61.37; 62.99; 64.67; 65.15; 68.87; 69.30; 70.15 (3×C);70.76; 70.92; 70.97; 73.03; 75.01; 75.33; 75.75; 76.84; 77.33; 77.89;99.96; 100.79; 100.86; 102.55; 103.80; 114.45 (d, 24.4 Hz); 117.62 (d,7.4 Hz); 119.92 (d, 7.5 Hz); 121.75 (d, 23.7 Hz); 157.82 (d, 174.2 Hz);170.33; 174.81; 175.52; 177.06; no resolution of the remaining signals.

EXAMPLE B12 Preparation of Compound No. (40)

(a) 31 mg (90%) of compound No. (42) are obtained according to ExampleB1.1(a1) (in this case the buffer solution comprises about 18% of DMSO(vol/vol)) from 26 mg (50 μmol) of compound No. (41) and 35 mg (57 μmol)of UDP-gal.

¹H-NMR (CD₃OD, 250.13 MHz) δ=0.40-1.41 (m, 12 H); 1.92 (t, 7.5 Hz, 2 H);3.32-4.01 (m, 17 H); 4.35 (d, 8.6 Hz, 1 H); 4.49 (d, 8.6 Hz, 1 H); 7.08(dd, 0.9 Hz, 7.6 Hz, 1 H); 7.33 (dd, 0.9 Hz, 7.6 Hz, 1 H); 7.45 (t, 7.6Hz, 1 H); 8.11 (d, 8.3 Hz, 1 H); 8.33 (d, 8.3 Hz, 1 H). ¹³C-NMR (CD₃OD,62.90 MHz) δ=25.78; 27.09; 29.92; 30.18; 30.25; 30.50; 34.65; 51.92;57.15; 62.04; 62.53; 70.32; 70.65; 72.63; 74.06; 74.83; 76.69; 77.15;81.24; 102.92; 105.15; 112.77; 118.98; 120.13; 130.58; 131.47; 138.45;138.83; 148.87; 155.06; 167.87; 175.89.

(b) 23 mg (80%) of compound No. (43) are obtained according to ExampleB2.1(c) (in this case the buffer solution comprises about 12% of DMSO(vol/vol)) from 20 mg (29 μmol) of compound No. (42) and 29 mg (44 μmol)of CMP-sia.

¹H-NMR (CD₃OD, 250.13 MHz) δ=0.41-1.41 (m, 12 H); 1.66 (broad t, 11.6Hz, 1 H); 1.95 (m, 5 H); 2.78 (dd, 11.6 Hz, 2.8 Hz, 1 H); 3.35-4.05 (m,24 H); 4.42 (d, 8.6 Hz, 1 H); 4.56 (d, 8.6 Hz, 1 H); 7.08 (dd, 0.9 Hz,7.6 Hz, 1 H); 7.32 (dd, 0.9 Hz, 7.6 Hz, 1 H); 7.45 (t, 7.6 Hz, 1 H);8.12 (d, 8.3 Hz, 1 H); 8.33 (d, 8.3 Hz, 1 H). ¹³C-NMR (CD₃OD, 62.90 MHz)δ=22.62; 25.78; 27.10; 29.74; 30.19; 30.25; 30.50; 34.66; 42.19; 51.92;53.95; 57.07; 62.11; 62.74; 64.49; 69.07; 69.32; 70.04; 70.67; 70.91;72.96; 74.10; 74.93; 76.68; 77.08; 77.66; 81.44; 101.12; 102.97; 105.08;112.89; 118.98; 120.05; 131.23; 131.54; 138.45; 138.88; 148.87; 155.06;167.57; 175.01; 175.51; 175.90.

(c) 5.0 mg (24%) of compound No. (40) are obtained according to ExampleB1.1(c) from 18.0 mg (18 μmol) of compound No. (43) and 17.2 mg (26μmol) of GDP-L-galactose. ¹H-NMR (CD₃OD, 400.13 MHz) δ=0.48-1.55 (m, 12H); 1.64 (broad t, 11.0 Hz, 1 H); 1.95 (s, 3 H); 1.97 (t, 7.6 Hz, 2 H);2.79 (dd, 11.6 Hz, 2.8 Hz, 1 H); 3.32-4.14 (m, 27 H); 4.50 (d, 8.6 Hz, 1H); 4.60 (m, 1 H); 4.68 (broad d, 8.6 Hz, 1 H); 5.13 (d, 4.3 Hz, 1 H);6.96 (dd, 0.9 Hz, 7.6 Hz, 1 H); 7.11 (dd, 0.9 Hz, 7.6 Hz, 1 H); 7.36 (t,7.6 Hz, 1 H); 8.00 (d, 8.3 Hz, 1 H); 8.20 (d, 8.3 Hz, 1 H). ¹³C-NMR(CD₃OD, 100.6 MHz) δ=22.56; 25.81; 27.10; 29.94; 30.19; 30.23; 30.54;34.70; 42.34; 53.96; 61.23; 62.44; 62.73; 64.70; 69.00; 69.33; 70.05;70.16; 70.73; 70.95 (3×C); 71.05 (2×C); 73.06; 75.02; 75.79; 76.82;77.45; 77.68; 99.96; 100.93; 102.86; 104.10; 113.81; 119.73; 132.08;138.49; no resolution of the remaining signals.

EXAMPLE B13 Preparation of Compound No. (44)

(a) 66 mg (89%) of compound No. (46) are obtained according to ExampleB1.1(a1) (in this case the buffer solution comprises about 7% of DMSO(vol/vol)) from 57 mg (111 μmol) of compound No. (45) and 92 mg (144μmol) of UDP-gal.

¹H-NMR (CD₃OD, 250.13 MHz) δ=1.02 (m, 8 H); 1.37 (m, 4 H); 2.11 (t, 7.5Hz, 2 H); 3.31-3.92 (m, 17 H); 4.36 (d, 8.6 Hz, 1 H); 4.50 (d, 8.6 Hz, 1H); 7.30 (dd, 2.1 Hz, 8.3 Hz, 1 H); 7.47 (d, 2.1 Hz, 1 H); 8.02 (d, 8.3Hz, 1 H). ¹³C-NMR (CD₃OD, 62.90 MHz) δ=25.71; 26.84; 29.78; 30.00 (2×C);30.28; 34.68; 52.29; 57.37; 61.14; 62.38; 70.20; 71.09; 72.45; 73.38;74.58; 76.44; 77.12; 80.55; 102.60; 104.79; 119.30; 120.77; 137.92;143.49; 153.28; 168.75; 176.72.

(b) 11 mg (27%) of compound No. (47) are obtained according to ExampleB1.1(b) (in this case the buffer solution comprises 9% of DMSO(vol/vol)) from 28 mg (42 μmol) of compound No. (46) and 40 mg (60 μmol)of CMP-sia.

¹H-NMR (CD₃OD, 250.13 MHz) δ=1.08 (m, 8 H); 1.39 (m, 4 H); 1.65 (broadt, 11.6 Hz, 1 H); 1.93 (s, 3 H); 2.13 (t, 7.6 Hz, 2 H); 2.78 (dd, 11.6Hz, 2.8 Hz, 1 H); 3.32-4.02 (m, 24 H); 4.40 (d, 8.6 Hz, 1 H); 4.46 (d,8.6 Hz, 1 H); 7.21 (dd, 2.1 Hz, 8.3 Hz, 1 H); 7.42 (d, 2.1 Hz, 1 H);8.00 (d, 8.3 Hz, 1 H). ¹³C-NMR (CD₃OD, 62.90 MHz) δ=22.60; 25.99; 27.17;30.07; 30.30; 30.38; 30.62; 34.71; 42.33; 51.97; 53.94; 57.23; 62.02;62.77; 64.49; 69.05; 69.33; 70.04; 70.70; 70.89; 72.95; 73.93; 74.94;76.59; 77.11; 77.65; 81.29; 101.51; 102.79; 105.00; 118.27; 121.53;126.66; 143.20; 168.55; 174.73; 175.63; 175.99; no resolution of theremaining signals.

(c) 6 mg (77%) of compound No. (44) are obtained according to ExampleA3(b2) from 7 mg (7 μmol) of compound No. (47) and 8 mg (12 μmol) ofGDP-L-gal. ¹H-NMR (CD₃OD, 400.13 MHz) δ=1.15 (m, 8 H); 1.43 (m, 4 H);1.65 (broad t, 11.0 Hz, 1 H); 1.96 (s, 3 H); 2.17 (t, 7.6 Hz, 2 H); 2.84(dd, 11.6 Hz, 2.8 Hz, 1 H); 3.25-4.11 (m, 27 H); 4.51 (d, 8.6 Hz, 1 H);4.54 (broad d, 8.6 Hz, 1 H); 4.66 (t, 6.4 Hz, 1 H); 5.04 (d, 4.3 Hz, 1H); 6.99 (dd, 2.1 Hz, 8.3 Hz, 1 H); 7.31 (d, 2.1 Hz, 1 H); 7.91 (d, 8.3Hz, 1 H). ¹³C-NMR (CD₃OD, 100.60 MHz) δ=22.57; 25.99; 27.17; 30.07;30.31 (2×C); 30.66; 34.76; 42.36; 51.95; 53.97; 58.18; 61.20; 62.37;62.72; 64.70; 67.70; 69.01; 69.31; 70.10; 70.77; 70.91; 71.00; 71.13;73.06; 75.02; 75.89; 76.64; 76.80; 77.37; 77.67; 100.22; 100.91; 102.39;104.11; 115.07; 123.11; 126.95; 139.41; 142.31; 169.62; 170.32; 174.74;175.51; 176.06.

C LIGAND BINDING ASSAY FOR DETERMINATION OF IC₅₀ VALUES—CONSERVED USE OFPOSITIVE CONTROLS

E-selectin/human IgG chimera [cloned and expressed according toKolbinger, F., Patton, J. T., Geisenhoff, G., Aenis, A., Li, X.,Katopodis, A., Biochemistry 35:6385-6392 (1996)] are incubated in Falconprobind™ microtiter plate (Plate 1) at a concentration of 200 ng/well in0.01 M Tris, 0.15 M NaCl, 1 mM CaCl₂, pH 7.4 (Tris-Ca⁺⁺ buffer). Thusthe plating solution is dispensed as 100 μl/well of 2 μg/ml E-chimera.Row 12 is left blank with only buffer. Plate 1 is incubated covered at37° C. for 2 hours. After incubation 100 μ/well of 2% BSA in Tris-Ca⁺⁺buffer is added and incubated at RT for 1 hour. During incubation thecompounds (2×serial dilution) are titrated in 1% BSA in Tris-Ca⁺⁺ usingU-shaped low bind microtiter plates (Plate 2). The rows are seriallydiluted up to row 9. Rows 10, 11, and 12 are just buffer. Final volumeis 60 μl/well and the first well contains 10 mM of compound with theexception of the positive controls, A (SLe^(x)-Lemieux) and B are usedas positive controls for each plate and the first well contains 5 mM ofthese compounds. PolySLe^(a)SA-HRP conjugate is prepared in advance byincubating Sialyl Le^(a)-PAA-biotin (cat #01-044, GlycoTech Corp.,Rockville, Md.) with Streptavidin-HRP in a molar ratio of 1:2. 60μl/well of 1 ng/μl of polySLe^(a)SA-HRP conjugate in 1% BSA in Tris-Ca⁺⁺are added to all wells except row 11 in Plate 2. Plate 1 is washed fourtimes with Tris-Ca⁺⁺ in the automatic plate washer. 100 μl/well aretransferred from Plate 2 to Plate 1 starting from lowest concentrationof compound. Plate 2 is discarded. The plate is incubated while rockingat RT for 2 hours. The plate is washed 4 times with Tris-Ca⁺⁺ usingautomatic plate washer. 100 μl/well of Substrate [Mix3,3′,5,5′-tetramethylbenzidine reagent and H₂O₂, at 1:1 ratio] are addedwith an 8 channel pipettor from right to left. The plate is incubated atRT for 2 minutes. The reaction is stopped by adding 100 μl/well of 1MH₃PO₄ using the 8 channel pipettor from right to left. Absorbance oflight at 450 nm is measured in a microtiter plate reader.

IC₅₀ is calculated by determining the concentration of compound requiredto inhibit maximal binding of the polySialylLe^(a)HRP conjugate toimmobilized E-selectin/human IgG chimera by 50%. The relative IC₅₀ iscalculated by determining the ratio of the IC₅₀ of an internal controlcompound to the IC₅₀ of the test compound.

In the following table RIC₅₀ means$\frac{{IC}_{50}( {{Test}\quad {compound}} )}{{IC}_{50}( {{Control}\quad {compound}\quad A} )}$

TABLE 1 Compound No. RIC₅₀ Compound No. RIC₅₀  (1) 2.780 (32) 5.602  (8)0.770 (36) 1.749 (10) 0.570 (37) 1.264 (14) 0.720 (40) 0.58 (21) 0.085(44) 0.246 (22) 1.137 (48) 0.376 (26) 0.366 (64) 1.066 (27) 0.711 (77)0.581 (31) 0.116

What is claimed is:
 1. A compound of the formula I or II

in which Z is an α-bonded pyranose of the formula III

 with the proviso that Z is not L-fucose, R₁ is hydrogen, C₁-C₂₀alkyl,C₁-C₂₀alkenyl, C₃-C₁₅cycloalkyl or a mono- or bicyclic C₆-C₁₀aryl orC₂-C₉heteroaryl, where alkyl, alkenyl, cycloalkyl, aryl and heteroarylare unsubstituted or mono- or polysubstituted by a substituent chosenfrom the group consisting of OH, halogen, halo-C₁-C₁₈alkyl, nitro,C₁-C₁₈alkyl, C₁-C₁₈alkoxy, amino, mono-C₁-C₁₈alkylamino,di-C₁-C₁₈alkylamino, benzylamino, sulfhydryl, thio-C₁-C₁₈alkyl andC₁-C₁₈alkylcarboxamide; R₂ is C₁-C₁₈alkyl, mono- or polysubstitutedC₁-C₁₈alkyl, C₃-C₈cycloalkyl or mono- or polysustituted C₃-C₈cycloalkyl,where one or more CH₂ groups in the alkyl and in the cycloalkyl,independently of one another may be replaced by oxygen, sulfur or animino group and the substituents are chosen from the group consisting ofOH, SH, NH₂, carboxamide, C(O)O and C₁-C₁₈alkoxycarbonyl; R₃ is a methylor hydroxymethyl group.
 2. A compound according to claim 1, in which R₁is hydrogen, C₁-C₂₀alkyl or C₁-C₂₀alkenyl, which are unsubstituted ormono- or polysubstituted by a substituent chosen from the groupconsisting of OH, halogen, halo-C₁-C₁₈alkyl, nitro, C₁-C₁₈alkyl,C₁-C₁₈alkoxy, amino, mono-C₁-C₁₈alkylamino, di-C₁-C₁₈alkylamino,benzylamino, sulfhydryl, thio-C₁-C₁₈alkyl and C₁-C₁₈alkylcarboxamide. 3.A compound according to claim 2, in which R₁ is C₁-C₁₀alkyl orC₁-C₁₀alkenyl, which are unsubstituted or mono- or polysubstituted by asubstituent chosen from the group consisting of OH, halogen,halo-C₁-C₁₈alkyl, nitro, C₁-C₁₈alkyl, C₁-C₁₈alkoxy, amino,mono-C₁-C₁₈alkylamino, di-C₁-C₁₈alkylamino, benzylamino, sulfhydryl,thio-C₁-C₁₈alkyl and C₁-C₁₈alkylcarboxamide.
 4. A compound according toclaim 3, in which R₁ is C₁-C₅alkyl or C₁-C₅alkenyl, which areunsubstituted or substituted by OH or halogen.
 5. A compound accordingto claim 4, in which R₁ is —CH₃, —CF₃, —CH₂—CH═CH₂, —CH₂OH or —CH₂SH. 6.A compound according to claim 1, in which R₁ is a mono- or bicyclicC₆-C₁₀aryl or C₂-C₉heteroaryl, which are unsubstituted or mono- orpolysubstituted by a substituent chosen from the group consisting of OH,halogen, halo-C₁-C₁₈alkyl, nitro, C₁-C₁₈alkyl, C₁-C₁₈alkoxy, amino,mono-C₁-C₁₈alkylamino, di-C₁-C₁₈alkylamino, benzylamino, sulfhydryl,thio-C₁-C₁₈alkyl and C₁-C₁₈alkylcarboxamide.
 7. A compound according toclaim 6, in which R₁ is a mono- or bicyclic C₆-C₁₀aryl orC₂-C₉heteroaryl, which are substituted by at least one OH and are notfurther substituted or are further mono- or polysubstituted by asubstituent chosen from the group consisting of halogen,halo-C₁-C₁₈alkyl, nitro, C₁-C₁₈alkyl, C₁-C₁₈alkoxy, amino,mono-C₁-C₁₈alkylamino, di-C₁-C₁₈alkylamino, benzylamino, sulfhydryl,thio-C₁-C₁₈alkyl and C₁-C₁₈alkylcarboxamide.
 8. A compound according toclaim 7, in which R₁ is phenyl or a mono- or bicyclic C₄-C₉heteroaryl,which are substituted by at least one OH and are not further substitutedor are further substituted by a substituent chosen from the groupconsisting of halogen, nitro, C₁-C₁₈alkyl and C₁-C₁₈alkoxy.
 9. Acompound according to claim 8, in which R₁ is phenyl, which issubstituted by one OH and F, NO₂, CH₃ or OCH₃ or by two OH; or in whichR₁ is a C₄heteroaryl which is substituted by two OH, or a C₉heteroarylwhich is substituted by one OH.
 10. A compound according to claim 1, inwhich R₂ is C₁-C₁₈alkyl, mono- or polysubstituted C₁-C₁₈alkyl,C₃-C₈cycloalkyl or mono- or polysubstituted C₃-C₈cycloalkyl, where thesubstituents are chosen from the group consisting of OH, SH, NH₂,carboxamide, C(O)O and C₁-C₁₈alkoxycarbonyl.
 11. A compound according toclaim 10, in which R₂ is C₁-C₁₈alkyl or C₁-C₁₈alkyl which is mono- orpolysubstituted independently of one another by OH, SH, NH₂,carboxamide, C(O)O or C₁-C₁₈alkoxycarbonyl.
 12. A compound according toclaim 11, in which R₂ is C₁-C₁₈alkyl or C₁-C₁₈alkyl which ismonosubstituted by C(O)O.
 13. A compound according to claim 12, in whichR₂ is —(CH₂)₈COOCH₃.
 14. A compound according to claim 1, in which R₃ ismethyl.
 15. A compound according to claim 1, in which the individual R₄independently of one another are hydrogen, OH, C₁-C₄alkyl, O-C₁-C₄alkyl,halogen, NH₂ or NHC(O)-C₁-C₈alkyl.
 16. A compound according to claim 15,in which the individual R₄ independently of one another are OH, halogenor NH₂.
 17. A compound according to claim 16, in which all the R₄ are OHor two R₄ are OH and one R₄ is halogen or NH₂.
 18. A compound accordingto claim 1, in which R₅ is hydrogen, C₁-C₈alkyl or (CH₂)_(m)OH, in whichm is a number from 1 to
 5. 19. A compound according to claim 18, inwhich R₅ is H, C₁-C₄alkyl or (CH₂)_(m)OH, in which m is 1 or
 2. 20. Acompound according to claim 19, in which R₅ is hydrogen, CH₃ or CH₂OH.21. A compound according to claim 1, in which X is —C(O)—, —S(O)₂— or—C(O)Y—, in which Y is —NH—, —S-C₁-C₆alkylene or —O-C₁-C₆alkylene.
 22. Acompound according to claim 21, in which X is —C(O)—, —S(O)₂—, —C(O)SCH₂or —C(O)OCH₂.
 23. A compound according to claim 1, in which R₁ ishydrogen, C₁-C₂₀alkyl or C₁-C₂₀alkenyl, which are unsubstituted or mono-or polysubstituted by a substituent chosen from the group consisting ofOH, halogen, halo-C₁-C₁₈alkyl, nitro, C₁-C₁₈alkyl, C₁-C₁₈alkoxy, amino,mono-C₁-C₁₈alkylamino, di-C₁-C₁₈alkylamino, benzylamino, sulfhydryl,thio-C₁-C₁₈alkyl and C₁-C₁₈alkylcarboxamide; R₂ is C₁-C₁₈alkyl, mono- orpolysubstituted C₁-C₁₈alkyl, C₃-C₈cycloalkyl or mono- or polysubstitutedC₃-C₈cycloalkyl, where the substituents are chosen from the groupconsisting of OH, SH, NH₂, carboxamide, C(O)O and C₁-C₁₈alkoxycarbonyl;R₃ is methyl; the individual R₄ independently of one another arehydrogen, OH, C₁-C₄alkyl, O-C₁-C₄alkyl, halogen, NH₂ orNHC(O)-C₁-C₈alkyl; R₅ is hydrogen, C₁-C₈alkyl or (CH₂)_(m)OH, in which mis a number from 1 to 5; and X is —C(O)—, —S(O)₂— or —C(O)Y—, in which Yis —NH—, —S-C₁-C₆alkylene or —O-C₁-C₆alkylene.
 24. A compound accordingto claim 23, in which R₁ is C₁-C₁₀alkyl or C₁-C₁₀alkenyl, which areunsubstituted or mono- or polysubstituted by a substituent chosen fromthe group consisting of OH, halogen, halo-C₁-C₁₈alkyl, nitro,C₁-C₁₈alkyl, C₁-C₁₈alkoxy, amino, mono-C₁-C₁₈alkylamino,di-C₁-C₁₈alkylamino, benzylamino, sulfhydryl, thio-C₁-C₁₈alkyl andC₁-C₁₈alkylcarboxamide; R₂ is C₁-C₁₈alkyl or C₁-C₁₈alkyl which is mono-or polysubstituted independently of one another by OH, SH, NH₂,carboxamide, C(O)O or C₁-C₁₈alkoxycarbonyl; R₃ is methyl; the individualR₄ independently of one another are OH, halogen or NH₂; R₅ is H,C₁-C₄alkyl or (CH₂)_(m)OH, in which m is 1 or 2; and X is —C(O)—,—S(O)₂—, —C(O)SCH₂ or —C(O)OCH₂.
 25. A compound according to claim 24,in which R₁ is C₁-C₅alkyl or C₁-C₅alkenyl, which are unsubstituted orsubstituted by OH or halogen; R₂ is C₁-C₁₈alkyl or C₁-C₁₈alkyl which ismonosubstituted by C(O)O; all the R₄ are OH or two R₄ are OH and one R₄is halogen or NH₂; and R₅ is hydrogen, CH₃ or CH₂OH.
 26. A compoundaccording to claim 25, in which R₁ is —CH₃, —CF₃, —CH₂—CH═CH₂, or—CH₂OH; R₂ is —(CH₂)₈COOCH₃; all the R₄ are OH or two R₄ are OH and oneR₄ is F or NH₂; and R₅ is hydrogen, CH₃ or CH₂OH.
 27. A compoundaccording to claim 1, in which R₁ is a mono- or bicyclic C₆-C₁₀aryl orC₂-C₉heteroaryl, which are unsubstituted or mono- or polysubstituted bya substituent chosen from the group consisting of OH, halogen,halo-C₁-C₁₈alkyl, nitro, C₁-C₁₈alkyl, C₁-C₁₈alkoxy, amino,mono-C₁-C₁₈alkylamino, di-C₁-C₁₈alkylamino, benzylamino, sulfhydryl,thio-C₁-C₁₈alkyl and C₁-C₁₈alkylcarboxamide; R₂ is C₁-C₁₈alkyl, mono- orpolysubstituted C₁-C₁₈alkyl, C₃-C₈cycloalkyl or mono- or polysubstitutedC₃-C₈cycloalkyl, where the substituents are chosen from the groupconsisting of OH, SH, NH₂, carboxamide, C(O)O and C₁-C₁₈alkoxycarbonyl;R₃ is methyl; the individual R₄ independently of one another arehydrogen, OH, C₁-C₄alkyl, O-C₁-C₄alkyl, halogen, NH₂ orNHC(O)-C₁-C₈alkyl; R₅ is hydrogen, C₁-C₈alkyl or (CH₂)_(m)OH, in which mis a number from 1 to 5; and X is —C(O)—, —S(O)₂— or —C(O)Y—, in which Yis —NH—, —S-C₁-C₆alkylene or —O-C₁-C₆alkylene.
 28. A compound accordingto claim 27, in which R₁ is a mono- or bicyclic C₆-C₁₈aryl orC₂-C₉heteroaryl, which are substituted by at least one OH and are notfurther substituted or are further mono- or polysubstituted by asubstituent chosen from the group consisting of halogen,halo-C₁-C₁₈alkyl, nitro, C₁-C₁₈alkyl, C₁-C₁₈alkoxy, amino,mono-C₁-C₁₈alkylamino, di-C₁-C₁₈alkylamino, benzylamino, sulfhydryl,thio-C₁-C₁₈alkyl and C₁-C₁₈alkylcarboxamide; R₂ is C₁-C₁₈alkyl orC₁-C₁₈alkyl which is mono- or polysubstituted independently of oneanother by OH, SH, NH₂, carboxamide, C(O)O or C₁-C₁₈alkoxycarbonyl; R₃is methyl; the individual R₄ independently of one another are OH,halogen or NH₂; R₅ is H, C₁-C₄alkyl or (CH₂)_(m)OH, in which m is 1 or2; and X is —C(O)—, —S(O)₂—, —C(O)SCH₂ or —C(O)OCH₂.
 29. A compoundaccording to claim 28, in which R₁ is phenyl or a mono- or bicyclicC₄-C₉heteroaryl, which are substituted by at least one OH and are notfurther substituted or are further substituted by a substituent chosenfrom the group consisting of halogen, nitro, C₁-C₁₈alkyl andC₁-C₁₈alkoxy; R₂ is C₁-C₁₈alkyl or C₁-C₁₈alkyl which is monosubstitutedby C(O)O; all the R₄ are OH or two R₄ are OH and one R₄ is halogen orNH₂; and R₅ is hydrogen, CH₃ or CH₂OH.
 30. A compound according to claim29, in which R₁ is phenyl, which is substituted by one OH and F, NO₂,CH₃ or OCH₃ or by two OH; or in which R₁ is a C₄heteroaryl which issubstituted by two OH, or a C₉heteroaryl which is substituted by one OH;R₂ is —(CH₂)₈COOCH₃; all the R₄ are OH or two R₄ are OH and one R₄ is For NH₂; and R₅ is hydrogen, CH₃ or CH₂OH.
 31. A compound of the formulaI according to claim 1, in which R₂ is —(CH₂)₈COOCH₃; R₃ is methyl; and(a) R₁ is hydrogen; Z is an α-bonded L-galactose; and X is —C(O)—; (b)R₁ is —CH₂—CH═CH₂; Z is an α-bonded L-galactose; and X is —C(O)OCH₂—;(c) R₁ is —CH₂—CH═CH₂; Z is an α-bonded D-arabinose; and X is—C(O)OCH₂—; (d) R₁ is 4-hydroxy-3-methoxy-phenyl; Z is an α-bondedD-arabinose; and X is —C(O)—; (e) R₁ is 4-hydroxy-3-methoxy-phenyl; Z isan α-bonded L-galactose, and X is —C(O)—; (f) R₁ is2-hydroxy-5-methyl-phenyl; Z is an α-bonded D-arabinose; and X is—C(O)—; (g) R₁ is 2-hydroxy-5-methyl-phenyl; Z is an α-bondedL-galactose; and X is —C(O)—; (h) R₁ is 2-hydroxy-3-nitro-phenyl; Z isan α-bonded L-galactose; and X is —C(O)—; (i) R₁ is2-hydroxy-5-fluoro-phenyl; Z is an α-bonded D-arabinose; and X is—C(O)—; (j) R₁ is 3,5-dihydroxy-phenyl; Z is an α-bonded D-arabinose;and X is —C(O)—; (k) R₁ is 3,5-dihydroxy-phenyl; Z is an α-bondedL-galactose; and X is —C(O)—; (l) R₁ is 3,5-dihydroxy-pyrimidinyl; Z isan α-bonded D-arabinose; and X is —C(O)—; (m) R₁ is3,5-dihydroxy-pyrimidinyl; Z is an α-bonded L-galactose; and X is—C(O)—; or (n) R₁ is 2-(8-hydroxy)quinolinyl; Z is an α-bondedL-galactose; and X is —C(O)—.
 32. A compound according to claim 31, inwhich R₂ is —(CH₂)₈COOCH₃; R₃ is methyl; Z is an α-bonded L-galactose; Xis —C(O)— and R₁ is hydrogen; 4-hydroxy-3-methoxy-phenyl;2-hydroxy-5-methyl-phenyl; 2-hydroxy-3-nitro-phenyl;3,5-dihydroxy-phenyl; 3,5-dihydroxy-pyrimidinyl or2-(8-hydroxy)quinolinyl.
 33. A compound according to claim 32, in whichR₂ is —(CH₂)₈COOCH₃; R₃ is methyl; Z is an α-bonded L-galactose; X is—C(O)— and R₁ is 4-hydroxy-3-methoxy-phenyl.
 34. A compound of theformula II according to claim 1, in which R₂ is —(CH₂)₈COOCH₃; R₃ ismethyl; and (a) R₁ is hydrogen; Z is an α-bonded D-arabinose; and X is—C(O)—; (b) R₁ is hydrogen; Z is an α-bonded L-2-fluoro-fucose; and X is—C(O)—; (c) R₁ is CH₃; Z is an α-bonded D-arabinose; and X is —C(O)—;(d) R₁ is CH₃; Z is an α-bonded L-2-fluoro-fucose; and X is —C(O)—; (e)R₁ is CH₃; Z is an α-bonded L-2-amino-fucose; and X is —C(O)—; (f) R₁ isCH₃; Z is an α-bonded L-galactose; and X is —C(O)—; (g) R₁ is CH₃; Z isan α-bonded L-glucose; and X is —C(O)—; (h) R₁ is CH₃; Z is an α-bondedL-galactose; and X is —C(O)OCH₂—; (i) R₁ is CH₃; Z is an α-bondedL-glucose; and X is —C(O)OCH₂—; (j) R₁ is CH₃; Z is an α-bondedD-arabinose; and X is S(O)₂; (k) R₁ is CH₃; Z is an α-bondedD-arabinose; and X is —C(O)SCH₂—; (l) R₁ is CF₃; Z is an α-bondedD-arabinose; and X is —C(O)—; (m) R₁ is CH₂OH; Z is an α-bondedD-arabinose; and X is —C(O)—; (n) R₁ is —CH₂—CH═CH₂; Z is an α-bondedD-arabinose; and X is —C(O)OCH₂—; (o) R₁ is —CH₂—CH═CH₂; Z is anα-bonded L-galactose; and X is —C(O)OCH₂—; (p) R₁ is phenyl; Z is anα-bonded L-galactose; and X is —C(O)OCH₂—; (q) R₁ is2-hydroxy-5-methyl-phenyl; Z is an α-bonded D-arabinose; and X is—C(O)—; (r) R₁ is 2-hydroxy-5-methyl-phenyl; Z is an α-bondedL-galactose; and X is —C(O)—; (s) R₁ is 2-hydroxy-5-fluoro-phenyl; Z isan α-bonded D-arabinose; and X is —C(O)—; (t) R₁ is4-hydroxy-3-methoxy-phenyl; Z is an α-bonded D-arabinose; and X is—C(O)—; (u) R₁ is 3,5-dihydroxy-phenyl; Z is an α-bonded L-galactose;and X is —C(O)—; (v) R₁ is 3,5-dihydroxy-phenyl; Z is an α-bondedL-2-amino-fucose; and X is —C(O)—; (w) R₁ is 3,5-dihydroxy-phenyl; Z isan α-bonded D-arabinose; and X is —C(O)OCH₂— or (x) R₁ is3,5-dihydroxy-pyrimidinyl; Z is an α-bonded D-arabinose; and X is—C(O)—.
 35. A compound according to claim 34, in which R₁ is CH₃; R₂ is—(CH₂)₈COOCH₃; R₃ is methyl; Z is an α-bonded L-galactose and X is—C(O)— or —C(O)OCH₂—.
 36. A process for the preparation of a compound ofthe formula I

in which Z is an α-bonded pyranose of the formula III

with the proviso that Z is not L-fucose, R₁ is hydrogen, C₁-C₂₀alkyl,C₁-C₂₀alkenyl, C₃-C₁₅cycloalkyl or a mono- or bicyclic C₆-C₁₀aryl orC₂-C₉heteroaryl, where alkyl, alkenyl, cycloalkyl, aryl and heteroarylare unsubstituted or mono- or polysubstituted by a substituent chosenfrom the group consisting of OH, halogen, halo-C₁-C₁₈alkyl, nitro,C₁-C₁₈alkyl, C₁-C₁₈alkoxy, amino, mono-C₁-C₁₈alkylamino,di-C₁-C₁₈alkylamino, benzylamino, sulfhydryl, thio-C₁-C₁₈alkyl andC₁-C₁₈alkylcarboxamide; R₂ is C₁-C₁₈alkyl, mono- or polysubstitutedC₁-C₁₈alkyl, C₃-C₈cycloalkyl or mono- or polysubstitutedC₃-C₈cycloalkyl, where one or more CH₂ groups in the alkyl and in thecycloalkyl, independently of one another may be replaced by oxygen,sulfur or an imino group and the substituents are chosen from the groupconsisting of OH, SH, NH₂, carboxamide, C(O)O and C₁-C₁₈alkoxycarbonyl;R₃ is a methyl or hydroxymethyl group; the individual R₄ independentlyof one another are hydrogen, OH, C₁-C₈alkyl, O-C₁-C₈alkyl, halogen, NH₂,SH or NHC(O-C₁-C₈alkyl; R₅ is hydrogen, C₁-C₈alkyl or (CH₂)_(m)R₄, inwhich m is a number from 1 to 5; and X is —C(O)—, —C(S)—, —S(O)₂—,—C(O)Y— or —C(S)Y—, in which Y is NH, O, S, S-C₁-C₆alkylene,NHC₁-C₆alkylene or O-C₁-C₆alkylene, which comprises (a) reacting acompound of the formula V R₇—X′—R₁  (V),  in which (a′) R₇ is halogen,X′ is as defined above for X and R₁ is as defined above, or (a″) R₇ isC(O) or C(S), X′ is —N═ and R₁ is as defined above, or (a″′) R₇ is OH,X′ is as defined above for X and R₁ is as defined above, directly afterin situ activation, with a compound of the formula IV

 in which R₂ is as defined above and the individual R₄ independently ofone another are hydrogen, acetyl, propionyl, butyryl or benzoyl, anyacetyl, propionyl, butyryl or benzoyl groups present being split offwith a basic alcohol solution, to give a compound of the formula VI

 in which R₂, R₁ and X are as defined above; (b) reacting the compoundof the formula VI with uridine di-phosphate-galactose in the presence ofβ(1→4)galactose transferase and then with cytidine mono-phosphate-sialicacid in the presence of α(2→3)sialic acid transferase to give a compoundof the formula VII

 in which R₁, R₂, R₃ and X are as defined above, and (c) reacting theresulting product with a guanosine di-phosphate-activated donor of theformula XI

 in which R₄ and R₅ are as defined above, in the presence of fucosetransferase VI to give a compound of the formula I.
 37. The process forthe preparation of a compound of the formula I according to claim 36,which comprises (a) reacting a compound of the formula VI according toclaim 36 with uridine di-phosphate-galactose in the presence ofβ(1→4)galactose transferase and then with cytidine mono-phosphate-sialicacid in the presence of α(2→3)sialic acid transferase to give a compoundof the formula VII according to claim 36 and (b) reacting the resultingproduct with a compound of the formula XI according to claim 36 in thepresence of fucose transferase to give a compound of the formula I. 38.The process for the preparation of a compound of the formula II

in which Z is an α-bonded pyranose of the formula III

with the proviso that Z is not L-fucose, R₁ is hydrogen, C₁-C₂₀alkyl,C₁-C₂₀alkenyl, C₃-C₁₅cycloalkyl or a mono- or bicyclic C₆-C₁₀aryl orC₂-C₉heteroaryl, where alkyl, alkenyl, cycloalkyl, aryl and heteroarylare unsubstituted or mono- or polysubstituted by a substituent chosenfrom the group consisting of OH, halogen, halo-C₁-C₁₈alkyl, nitro,C₁-C₁₈alkyl, C₁-C₁₈alkoxy, amino, mono-C₁-C₁₈alkylamino,di-C₁-C₁₈alkylamino, benzylamino, sulfhydryl, thio-C₁-C₁₈alkyl andC₁-C₁₈alkylcarboxamide; R₂ is C₁-C₁₈alkyl, mono- or polysubstitutedC₁-C₁₈alkyl, C₃-C₈cycloalkyl or mono- or polysubstitutedC₃-C₈cycloalkyl, where one or more CH₂ groups in the alkyl and in thecycloalkyl, independently of one another may be replaced by oxygen,sulfur or an imino group and the substituents are chosen from the groupconsisting of OH, SH, NH₂, carboxamide, C(O)O and C₁-C₁₈alkoxycarbonyl;R₃ is a methyl or hydroxymethyl group; the individual R₄ independentlyof one another are hydrogen, OH, C₁-C₈alkyl, O-C₁-C₈alkyl, halogen, NH₂,SH or NHC(O-C₁-C₈alkyl; R₅ is hydrogen, C₁-C₈alkyl or (CH₂)_(m)R₄, inwhich m is a number from 1 to 5; and X is —C(O)—, —C(S)—, —S(O)₂—,—C(O)Y— or —C(S)Y—, in which Y is NH, O, S, S-C₁-C₆alkylene,NH-C₁-C₆alkylene or O-C₁-C₆alkylene, which comprises (a) reacting acompound of the formula IV with a compound of the formula V according toclaim 36, (b) reacting the compound of the formula VI according to claim36 with uridine di-phosphate-galactose in the presence ofβ(1→3)galactose transferase and then cytidine mono-phosphate-sialic acidin the presence of α(2→3)sialic acid transferase to give a compound ofthe formula VIII

 in which R₁, R₂, R₃ and X are as defined above, and (c) reacting theresulting product with a compound of the formula XI according to claim36 in the presence of fucose transferase to give a compound of theformula II.
 39. The process for the preparation of a compound of theformula II

which comprises (a) reacting a compound of the formula V R₇—X′—R₁  (V), in which (a′) R₇ is halogen, X′ is as defined for X according to claim36 and R₁ is as defined according to claim 36, or (a″) R₇ is C(O) orC(S), X′ is —N═ and R₁ is as defined above, or (a″′) R₇ is OH, X′ is asdefined above for X and R₁ is as defined above, directly after in situactivation, with a compound of the formula IX

 in which R₂ is as defined above and the individual R₄ independently ofone another are hydrogen, acetyl, propionyl, butyryl or benzoyl, anyacetyl, propionyl, butyryl or benzoyl groups present being split offwith a basic alcohol solution, to give a compound of the formula X

 in which R₂, R₁ and X are as defined above; (b) reacting the compoundof the formula X with cytidine mono-phosphate-sialic acid in thepresence of α(2→3)sialic acid transferase to give a compound of theformula VIII

 in which R₁, R₂, R₃ and X are as defined above, and (c) reacting theresulting product with a compound of the formula XI in the presence offucose transferase to give a compound of the formula II.
 40. The processfor the preparation of a compound of the formula II according to claim36, which comprises (a) reacting a compound of the formula X accordingto claim 36 with cytidine mono-phosphate-sialic acid in the presence ofα(2→3)sialic acid transferase to give a compound of the formula VIIIaccording to claim 36 and (b) reacting the resulting product with acompound of the formula XI according to claim 36 in the presence offucose transferase to give a compound of the formula II.
 41. The processfor the preparation of a compound of the formula II

which comprises (a) reacting a compound of the formula VI according toclaim 36 with uridine di-phosphate-galactose in the presence of β(1→3)galactose transferase and then with cytidine mono-phosphate-sialic acidin the presence of α(2→3) sialic acid transferase to give a compound ofthe formula VII

 in which R₁, R₂, R₃ and are as defined according to claim 36, and (b)reacting the resulting product with a compound of the formula XIaccording to claim 36 in the presence of fucose transferase to give acompound of the formula II.
 42. The process as claimed in claim 36,wherein the galactosylation and the sialylation are carried outsimultaneously.
 43. The process as claimed in claim 36, wherein thegalactosylation and the sialylation are carried out in succession. 44.The process as claimed in claim 37, wherein the galactosylation and thesialylation are carried out simultaneously.
 45. The process as claimedin claim 37, wherein the galactosylation and the sialylation are carriedout in succession.
 46. The process as claimed in claim 38, wherein thegalactosylation and the sialylation are carried out simultaneously. 47.The process as claimed in claim 38 wherein the galactosylation and thesialylation are carried out in succession.
 48. The process as claimed inclaim 39, wherein the galactosylation and the sialylation are carriedout simultaneously.
 49. The process as claimed in claim 39, wherein thegalactosylation and the sialylation are carried out in succession. 50.The process as claimed in claim 41, wherein the galactosylation and thesialylation are carried out simultaneously.
 51. The process as claimedin claim 41, wherein the galactosylation and the sialylation are carriedout in succession.
 52. A pharmaceutical preparation comprising an activeamount of a compound according to claim 1, by itself or together withother active ingredients, and a pharmaceutical carrier.
 53. Thepharmaceutical preparation according to claim 52 further including anadjunct.